Substituted biaryl piperazinyl-pyridine analogues

ABSTRACT

Substituted biaryl piperazinyl-pyridine analogues are provided, of the Formula: 
                         
wherein variables are as described herein. Such compounds are ligands that may be used to modulate specific receptor activity in vivo or in vitro, and are particularly useful in the treatment of conditions associated with pathological receptor activation in humans, domesticated companion animals and livestock animals. Pharmaceutical compositions and methods for using such compounds to treat such disorders are provided, as are methods for using such ligands for receptor localization studies.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.11/204,202, filed Aug. 13, 2005, now issued U.S. Pat. No. 7,662,830,which claims the benefit of U.S. Provisional Patent Application Nos.60/601,721, filed Aug. 13, 2004, and 60/641,796, filed Jan. 5, 2005. Thecontents of each of the foregoing applications are incorporated hereinby reference in their entirety.

FIELD OF THE INVENTION

This invention relates generally to substituted biarylpiperazinyl-pyridine analogues that have useful pharmacologicalproperties. The invention further relates to the use of such compoundsfor treating conditions related to capsaicin receptor activation, foridentifying other agents that bind to capsaicin receptor, and as probesfor the detection and localization of capsaicin receptors.

BACKGROUND OF THE INVENTION

Pain perception, or nociception, is mediated by the peripheral terminalsof a group of specialized sensory neurons, termed “nociceptors.” A widevariety of physical and chemical stimuli induce activation of suchneurons in mammals, leading to recognition of a potentially harmfulstimulus. Inappropriate or excessive activation of nociceptors, however,can result in debilitating acute or chronic pain.

Neuropathic pain involves pain signal transmission in the absence ofstimulus, and typically results from damage to the nervous system. Inmost instances, such pain is thought to occur because of sensitizationin the peripheral and central nervous systems following initial damageto the peripheral system (e.g., via direct injury or systemic disease).Neuropathic pain is typically burning, shooting and unrelenting in itsintensity and can sometimes be more debilitating that the initial injuryor disease process that induced it.

Existing treatments for neuropathic pain are largely ineffective.Opiates, such as morphine, are potent analgesics, but their usefulnessis limited because of adverse side effects, such as physicaladdictiveness and withdrawal properties, as well as respiratorydepression, mood changes, and decreased intestinal motility withconcomitant constipation, nausea, vomiting, and alterations in theendocrine and autonomic nervous systems. In addition, neuropathic painis frequently non-responsive or only partially responsive toconventional opioid analgesic regimens. Treatments employing theN-methyl-D-aspartate antagonist ketamine or the alpha(2)-adrenergicagonist clonidine can reduce acute or chronic pain, and permit areduction in opioid consumption, but these agents are often poorlytolerated due to side effects.

Topical treatment with capsaicin has been used to treat chronic andacute pain, including neuropathic pain. Capsaicin is a pungent substancederived from the plants of the Solanaceae family (which includes hotchili peppers) and appears to act selectively on the small diameterafferent nerve fibers (A-delta and C fibers) that are believed tomediate pain. The response to capsaicin is characterized by persistentactivation of nociceptors in peripheral tissues, followed by eventualdesensitization of peripheral nociceptors to one or more stimuli. Fromstudies in animals, capsaicin appears to trigger C fiber membranedepolarization by opening cation selective channels for calcium andsodium.

Similar responses are also evoked by structural analogues of capsaicinthat share a common vanilloid moiety. One such analogue isresiniferatoxin (RTX), a natural product of Euphorbia plants. The termvanilloid receptor (VR) was coined to describe the neuronal membranerecognition site for capsaicin and such related irritant compounds. Thecapsaicin response is competitively inhibited (and thereby antagonized)by another capsaicin analog, capsazepine, and is also inhibited by thenon-selective cation channel blocker ruthenium red, which binds to VRwith no more than moderate affinity (typically with a K_(i) value of nolower than 140 μM).

Rat and human vanilloid receptors have been cloned from dorsal rootganglion cells. The first type of vanilloid receptor to be identified isknown as vanilloid receptor type 1 (VR1), and the terms “VR1” and“capsaicin receptor” are used interchangeably herein to refer to ratand/or human receptors of this type, as well as mammalian homologues.The role of VR1 in pain sensation has been confirmed using mice lackingthis receptor, which exhibit no vanilloid-evoked pain behavior andimpaired responses to heat and inflammation. VR1 is a nonselectivecation channel with a threshold for opening that is lowered in responseto elevated temperatures, low pH, and capsaicin receptor agonists.Opening of the capsaicin receptor channel is generally followed by therelease of inflammatory peptides from neurons expressing the receptorand other nearby neurons, increasing the pain response. After initialactivation by capsaicin, the capsaicin receptor undergoes a rapiddesensitization via phosphorylation by cAMP-dependent protein kinase.

Because of their ability to desensitize nociceptors in peripheraltissues, VR1 agonist vanilloid compounds have been used as topicalanesthetics. However, agonist application may itself cause burning pain,which limits this therapeutic use. Recently, it has been reported thatVR1 antagonists, including certain nonvanilloid compounds, are alsouseful for the treatment of pain (see, e.g., PCT InternationalApplication Publication Numbers WO 02/08221, WO 03/062209, WO 04/054582,WO 04/055003, WO 04/055004, WO 04/056774, WO 05/007646, WO 05/007648, WO05/007652, WO 05/009977, WO 05/009980 and WO 05/009982).

Thus, compounds that interact with VR1, but do not elicit the initialpainful sensation of VR1 agonist vanilloid compounds, are desirable forthe treatment of chronic and acute pain, including neuropathic pain, aswell as other conditions that are responsive to capsaicin receptormodulation. The present invention fulfills this need, and providesfurther related advantages.

SUMMARY OF THE INVENTION

The present invention provides compounds of Formula I:

as well as pharmaceutically acceptable salts of such compounds. WithinFormula I:

-   Ar₁ is phenyl or a 6-membered aromatic heterocycle, each of which is    substituted with R₁₀ and with from 0 to 4 additional substituents    (e.g., independently chosen from R₁);-   Ar₂ is phenyl or a 6-membered aromatic heterocycle, each of which is    optionally substituted, (e.g., with from 0 to 4 substituents    independently chosen from R₂);-   X, Y and Z are independently optionally substituted carbon (e.g.,    CR_(x)) or N, such that at least one of X, Y and Z is N;-   R_(x), is independently chosen at each occurrence from hydrogen,    halogen, C₁-C₄alkyl, amino, cyano and mono- and    di-(C₁-C₄alkyl)amino;-   Each R₁ is independently chosen from:    -   (a) halogen, cyano and nitro;    -   (b) groups of the formula -Q-M-R_(y); and    -   (c) groups that are taken together with R₁₀ to form a fused 5-        to 7-membered carbocyclic or heterocyclic ring that is        optionally substituted (e.g., with from 0 to 4 substituents        independently chosen from halogen, cyano, nitro and groups of        the formula -Q-M-R_(y));-   R₁₀ represents one substituent chosen from:    -   (a) nitro;    -   (b) groups of the formula -Q-M-R_(y); and    -   (c) groups that are taken together with a substituent        represented by R₁ to form a fused, optionally substituted 5- to        7-membered carbocyclic or heterocyclic ring;    -   such that, in certain aspects, R₁₀ is not hydroxy, amino or an        unsubstituted group chosen from C₁-C₆alkyl, C₂-C₆alkenyl,        C₂-C₆alkynyl, C₁-C₆alkoxy, C₂-C₆alkyl ether, C₂-C₆alkanoyl,        C₃-C₆alkanone, C₁-C₆haloalkyl, C₁-C₆haloalkoxy, mono- or        di-(C₁-C₆alkyl)amino, C₁-C₆alkylsulfonyl, mono- or        di-(C₁-C₆alkyl)aminosulfonyl or mono- or        di-(C₁-C₆alkyl)aminocarbonyl;-   Each Q is independently chosen from C₀-C₄alkylene;-   Each M is independently absent or selected from O, C(═O)

-    wherein m is independently selected at each occurrence from 0, 1    and 2; and R_(z) is independently selected at each occurrence from    hydrogen, C₁-C₈alkyl and groups that are taken together with R_(y)    to form an optionally substituted 4- to 7-membered heterocycle;-   Each R_(y) is independently hydrogen, C₁-C₈haloalkyl, optionally    substituted C₁-C₈alkyl, optionally substituted C₂-C₈alkenyl,    optionally substituted (C₃-C₈-carbocycle)C₀-C₄alkyl, optionally    substituted (4- to 7-membered heterocycle)C₀-C₄alkyl, or taken    together with R_(z) to form an optionally substituted 4- to    7-membered heterocycle (e.g., in certain embodiments each alkyl,    carbocycle and heterocycle is substituted with from 0 to 4    substituents independently selected from hydroxy, halogen, amino,    cyano, nitro, oxo, —COOH, aminocarbonyl, aminosulfonyl, C₁-C₆alkyl,    C₃-C₇cycloalkyl, C₂-C₆alkyl ether, C₁-C₆alkanoyl,    C₁-C₆alkylsulfonyl, C₁-C₈alkoxy, C₁-C₈hydroxyalkyl, C₁-C₈ alkylthio,    mono- and di-(C₁-C₆alkyl)aminocarbonyl, C₁-C₆alkanoylamino, mono-    and di-(C₁-C₆alkyl)aminosulfonyl, mono- and di-(C₁-C₆alkyl)amino and    phenyl); such that R_(y) is not hydrogen if Q is a single covalent    bond (i.e., C₀alkyl) and M is absent;-   Each R₂ is:    -   (a) independently chosen from (i) hydroxy, amino, cyano,        halogen, —COOH, aminosulfonyl, nitro and aminocarbonyl; and (ii)        C₁-C₆alkyl, (C₃-C₈cycloalkyl)C₀-C₄alkyl, C₁-C₆haloalkyl,        C₁-C₆alkoxy, C₁-C₆alkylthio, C₂-C₆alkyl ether, C₂-C₆alkanoyl,        C₁-C₆alkoxycarbonyl, C₂-C₆alkanoyloxy, C₃-C₆alkanone, mono- and        di-(C₁-C₈alkyl)aminoC₀-C₆alkyl, (4- to 7-membered        heterocycle)C₀-C₄alkyl, C₁-C₆alkylsulfonyl, mono- and        di-(C₁-C₆alkyl)aminosulfonyl, and mono- and        di-(C₁-C₆alkyl)aminocarbonyl, each of which is optionally        substituted, (e.g., with from 0 to 4 substituents independently        chosen from halogen, hydroxy, cyano, amino, aminocarbonyl,        aminosulfonyl, —COOH and oxo); or    -   (b) taken together with an adjacent R₂ to form a fused 5- to        13-membered carbocyclic or heterocyclic group that is optionally        substituted (e.g., with from 0 to 3 substituents independently        chosen from halogen, oxo and C₁-C₆alkyl);-   R₃ is selected from:    -   (i) hydrogen and halogen;    -   (ii) C₁-C₆alkyl, (C₃-C₈cycloalkyl)C₀-C₂alkyl, C₁-C₆haloalkyl and        phenylC₀-C₂alkyl; and    -   (iii) groups of the formula:

-   -   -   wherein:        -   L is C₀-C₆alkylene or C₁-C₆alkylene that is taken together            with R₅, R₆ or R₇ to form a 4- to 7-membered heterocycle;        -   W is O, CO, S, SO or SO₂;        -   R₅ and R₆ are:            -   (a) independently chosen from hydrogen, C₁-C₁₂alkyl,                C₂-C₁₂alkenyl, (C₃-C₈cycloalkyl)C₀-C₄alkyl,                C₂-C₆alkanoyl, C₁-C₆alkylsulfonyl, phenylC₀-C₆alkyl, (4-                to 7-membered heterocycle)C₀-C₆alkyl and groups that are                joined to L to form a 4- to 7-membered heterocycle; or            -   (b) joined to form a 4- to 12-membered heterocycle; and        -   R₇ is hydrogen, C₁-C₁₂alkyl, C₂-C₁₂alkenyl,            (C₃-C₈cycloalkyl)C₀-C₄alkyl, C₂-C₆alkanoyl,            phenylC₀-C₆alkyl, (4- to 7-membered heterocycle)C₀-C₆alkyl            or a group that is joined to L to form a 4- to 7-membered            heterocycle;

    -   wherein each of (ii) and (iii) is optionally substituted (e.g.,        with from 0 to 4 substituents independently chosen from:        -   (1) halogen, hydroxy, amino, cyano, nitro, —COOH,            aminosulfonyl, aminocarbonyl and oxo; and        -   (2) C₁-C₆alkyl, (C₃-C₈cycloalkyl)C₀-C₂alkyl, C₁-C₆haloalkyl,            C₁-C₆alkoxy, C₁-C₆alkanoyl, C₁-C₆alkoxycarbonyl,            C₂-C₆alkanoylamino, mono- and            di-(C₁-C₆alkyl)aminoC₀-C₄alkyl, C₁-C₆alkylsulfonyl, mono-            and di-(C₁-C₆alkyl)aminosulfonyl, mono- and            di-(C₁-C₆alkyl)aminocarbonylC₀-C₄alkyl, phenylC₀-C₄alkyl and            (4- to 7-membered heterocycle)C₀-C₄alkyl, each of which is            substituted with from 0 to 4 secondary substituents            independently chosen from halogen, hydroxy, cyano, oxo,            imino, C₁-C₄alkyl, C₁-C₄alkoxy and C₁-C₄haloalkyl); and

-   R₄ represents from 0 to 2 substituents (e.g., substituents that are    independently chosen from C₁-C₃alkyl, C₁-C₃haloalkyl and oxo).

Within certain aspects, substituted biaryl piperazinyl-pyridineanalogues provided herein are VR1 modulators and exhibit a K_(i) of nogreater than 1 micromolar, 500 nanomolar, 100 nanomolar, 50 nanomolar,10 nanomolar or 1 nanomolar in a capsaicin receptor binding assay and/orhave an EC₅₀ or IC₅₀ value of no greater than 1 micromolar, 500nanomolar, 100 nanomolar, 50 nanomolar, 10 nanomolar or 1 nanomolar inan in vitro assay for determination of capsaicin receptor agonist orantagonist activity. In certain embodiments, such VR1 modulators are VR1antagonists and exhibit no detectable agonist activity in an in vitroassay of capsaicin receptor activation at a concentration equal to theIC₅₀, 10 times the IC₅₀ or 100 times the IC₅₀.

Within certain aspects, compounds as described herein are labeled with adetectable marker (e.g., radiolabeled or fluorescein conjugated).

The present invention further provides, within other aspects,pharmaceutical compositions comprising at least one substituted biarylpiperazinyl-pyridine analogue provided herein in combination with aphysiologically acceptable carrier or excipient.

Within further aspects, methods are provided for reducing calciumconductance of a cellular capsaicin receptor, comprising contacting acell (e.g., neuronal) expressing a capsaicin receptor with at least oneVR1 modulator as described herein. Such contact may occur in vivo or invitro.

Methods are further provided for inhibiting binding of vanilloid ligandto a capsaicin receptor. Within certain such aspects, the inhibitiontakes place in vitro. Such methods comprise contacting a capsaicinreceptor with at least one VR1 modulator as described herein, underconditions and in an amount or concentration sufficient to detectablyinhibit vanilloid ligand binding to the capsaicin receptor. Within othersuch aspects, the capsaicin receptor is in a patient. Such methodscomprise contacting cells expressing a capsaicin receptor in a patientwith at least one VR1 modulator as described herein in an amount orconcentration that would be sufficient to detectably inhibit vanilloidligand binding to cells expressing a cloned capsaicin receptor in vitro,and thereby inhibiting binding of vanilloid ligand to the capsaicinreceptor in the patient.

The present invention further provides methods for treating a conditionresponsive to capsaicin receptor modulation in a patient, comprisingadministering to the patient a therapeutically effective amount of atleast one VR1 modulator as described herein.

Within other aspects, methods are provided for treating pain in apatient, comprising administering to a patient suffering from pain atherapeutically effective amount of at least one substituted biarylpiperazinyl-pyridine analogue.

Methods are further provided for treating itch, urinary incontinence,overactive bladder, cough and/or hiccup in a patient, comprisingadministering to a patient suffering from one or more of the foregoingconditions a therapeutically effective amount of at least onesubstituted biaryl piperazinyl-pyridine analogue.

The present invention further provides methods for promoting weight lossin an obese patient, comprising administering to an obese patient atherapeutically effective amount of at least one substituted biarylpiperazinyl-pyridine analogue.

Methods are further provided for identifying an agent that binds tocapsaicin receptor, comprising: (a) contacting capsaicin receptor with alabeled compound as described herein under conditions that permitbinding of the compound to capsaicin receptor, thereby generating bound,labeled compound; (b) detecting a signal that corresponds to the amountof bound, labeled compound in the absence of test agent; (c) contactingthe bound, labeled compound with a test agent; (d) detecting a signalthat corresponds to the amount of bound labeled compound in the presenceof test agent; and (e) detecting a decrease in signal detected in step(d), as compared to the signal detected in step (b).

Within further aspects, the present invention provides methods fordetermining the presence or absence of capsaicin receptor in a sample,comprising: (a) contacting a sample with a compound as described hereinunder conditions that permit binding of the compound to capsaicinreceptor; and (b) detecting a signal indicative of a level of thecompound bound to capsaicin receptor.

The present invention also provides packaged pharmaceuticalpreparations, comprising: (a) a pharmaceutical composition as describedherein in a container; and (b) instructions for using the composition totreat one or more conditions responsive to capsaicin receptormodulation, such as pain, itch, urinary incontinence, overactivebladder, cough, hiccup and/or obesity.

In yet another aspect, the present invention provides methods forpreparing the compounds disclosed herein, including the intermediates.

These and other aspects of the invention will become apparent uponreference to the following detailed description.

DETAILED DESCRIPTION

As noted above, the present invention provides substituted biarylpiperazinyl-pyridine analogues. Such compounds may be used in vitro orin vivo, to modulate capsaicin receptor activity in a variety ofcontexts.

Terminology

Compounds are generally described herein using standard nomenclature.For compounds having asymmetric centers, it should be understood that(unless otherwise specified) all of the optical isomers and mixturesthereof are encompassed. In addition, compounds with carbon-carbondouble bonds may occur in Z- and E-forms, with all isomeric forms of thecompounds being included in the present invention unless otherwisespecified. Where a compound exists in various tautomeric forms, arecited compound is not limited to any one specific tautomer, but ratheris intended to encompass all tautomeric forms. Certain compounds aredescribed herein using a general formula that includes variables (e.g.,R₃, Ar₁, Z). Unless otherwise specified, each variable within such aformula is defined independently of any other variable, and any variablethat occurs more than one time in a formula is defined independently ateach occurrence.

The term “substituted biaryl piperazinyl-pyridine analogues,” as usedherein, encompasses all compounds of Formula I (including compounds ofother Formulas provided herein, as well as any enantiomers, racematesand stereoisomers) and pharmaceutically acceptable salts of suchcompounds. Compounds in which the core ring

is pyridyl, pyrimidyl or triazinyl (e.g.,

are specifically included within the definition of substituted biarylpiperazinyl-pyridine analogues.

A “pharmaceutically acceptable salt” of a compound recited herein is anacid or base salt that is generally considered in the art to be suitablefor use in contact with the tissues of human beings or animals withoutexcessive toxicity or carcinogenicity, and preferably withoutirritation, allergic response, or other problem or complication. Suchsalts include mineral and organic acid salts of basic residues such asamines, as well as alkali or organic salts of acidic residues such ascarboxylic acids. Specific pharmaceutical salts include, but are notlimited to, salts of acids such as hydrochloric, phosphoric, hydroponic,malic, glycolic, fumaric, sulfuric, sulfamic, sulfanilic, formic,toluenesulfonic, methanesulfonic, benzene sulfonic, ethane disulfonic,2-hydroxyethylsulfonic, nitric, benzoic, 2-acetoxybenzoic, citric,tartaric, lactic, stearic, salicylic, glutamic, ascorbic, pamoic,succinic, fumaric, maleic, propionic, hydroxymaleic, hydroiodic,phenylacetic, alkanoic such as acetic, HOOC—(CH₂)_(n)—COOH where n is0-4, and the like. Similarly, pharmaceutically acceptable cationsinclude, but are not limited to sodium, potassium, calcium, aluminum,lithium and ammonium. Those of ordinary skill in the art will recognizefurther pharmaceutically acceptable salts for the compounds providedherein, including those listed by Remington's Pharmaceutical Sciences,17th ed., Mack Publishing Company, Easton, Pa., p. 1418 (1985). Ingeneral, a pharmaceutically acceptable acid or base salt can besynthesized from a parent compound that contains a basic or acidicmoiety by any conventional chemical method. Briefly, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in water or inan organic solvent, or in a mixture of the two; generally, the use ofnonaqueous media, such as ether, ethyl acetate, ethanol, isopropanol oracetonitrile, is preferred.

It will be apparent that each compound of Formula I may, but need not,be formulated as a hydrate, solvate or non-covalent complex. Inaddition, the various crystal forms and polymorphs are within the scopeof the present invention, as are prodrugs of the compounds of Formula I.A “prodrug” is a compound that may not fully satisfy the structuralrequirements of the compounds provided herein, but is modified in vivo,following administration to a patient, to produce a compound of FormulaI, or other formula provided herein. For example, a prodrug may be anacylated derivative of a compound as provided herein. Prodrugs includecompounds wherein hydroxy, amine or sulfhydryl groups are bonded to anygroup that, when administered to a mammalian subject, cleaves to form afree hydroxy, amino, or sulfhydryl group, respectively. Examples ofprodrugs include, but are not limited to, acetate, formate, phosphateand benzoate derivatives of alcohol and amine functional groups withinthe compounds provided herein. Prodrugs of the compounds provided hereinmay be prepared by modifying functional groups present in the compoundsin such a way that the modifications are cleaved in vivo to yield theparent compounds.

As used herein (except when used in the terms “alkylamino” and“alkylaminoalkyl,” as discussed below), the term “alkyl” refers to astraight or branched chain saturated aliphatic hydrocarbon. Alkyl groupsinclude groups having from 1 to 8 carbon atoms (C₁-C₈alkyl), from 1 to 6carbon atoms (C₁-C₆alkyl) and from 1 to 4 carbon atoms (C₁-C₄alkyl),such as methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl,tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl,3-hexyl and 3-methylpentyl. “C₀-C_(n)alkyl” refers to a single covalentbond (C₀) or an alkyl group having from 1 to n carbon atoms; for example“C₀-C₄alkyl” refers to a single covalent bond or a C₁-C₄alkyl group;“C₀-C₈alkyl” refers to a single covalent bond or a C₁-C₈alkyl group. Insome instances, a substituent of an alkyl group is specificallyindicated. For example, “C₁-C₄hydroxyalkyl” refers to a C₁-C₄alkyl groupthat has at least one hydroxy substituent.

“Alkylene” refers to a divalent alkyl group, as defined above.C₀-C₄alkylene is a single covalent bond or an alkylene group having from1 to 4 carbon atoms; and C₀-C₆alkylene is a single covalent bond or analkylene group having from 1 to 6 carbon atoms.

“Alkenyl” refers to straight or branched chain alkene groups, whichcomprise at least one unsaturated carbon-carbon double bond. Alkenylgroups include C₂-C₈alkenyl, C₂-C₆alkenyl and C₂-C₄alkenyl groups, whichhave from 2 to 8, 2 to 6 or 2 to 4 carbon atoms, respectively, such asethenyl, allyl or isopropenyl. “Alkynyl” refers to straight or branchedchain alkyne groups, which have one or more unsaturated carbon-carbonbonds, at least one of which is a triple bond. Alkynyl groups includeC₂-C₈alkynyl, C₂-C₆alkynyl and C₂-C₄alkynyl groups, which have from 2 to8, 2 to 6 or 2 to 4 carbon atoms, respectively.

A “cycloalkyl” is a group that comprises one or more saturated and/orpartially saturated rings in which all ring members are carbon, such ascyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,cyclooctyl, adamantyl, decahydro-naphthalenyl, octahydro-indenyl, andpartially saturated variants of the foregoing, such as cyclohexenyl.Cycloalkyl groups do not comprise an aromatic ring or a heterocyclicring. Certain cycloalkyl groups are C₃-C₈cycloalkyl, in which the groupcontains a single ring with from 3 to 8 ring members. A“(C₃-C₈cycloalkyl)C₀-C₄alkyl” is a C₃-C₈cycloalkyl group linked via asingle covalent bond or a C₁-C₄alkylene group.

By “alkoxy,” as used herein, is meant an alkyl group as described aboveattached via an oxygen bridge. Alkoxy groups include C₁-C₆alkoxy andC₁-C₄alkoxy groups, which have from 1 to 6 or from 1 to 4 carbon atoms,respectively. Methoxy, ethoxy, propoxy, isopropoxy, n-butoxy,sec-butoxy, tert-butoxy, n-pentoxy, 2-pentoxy, 3-pentoxy, isopentoxy,neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, and 3-methylpentoxy arerepresentative alkoxy groups. Similarly, “alkylthio” refers to an alkylgroup as described above attached via a sulfur bridge.

The term “oxo,” as used herein, refers to a keto group (C═O). An oxogroup that is a substituent of a nonaromatic carbon atom results in aconversion of —CH₂— to —C(═O)—.

Similarly, an “imino” is a group of the formula C═N. The term“iminoalkyl” refers to an alkyl group as described above substitutedwith an imine (e.g., a group of the formula

The term “alkanoyl” refers to an acyl group (e.g., —(C═O)-alkyl), inwhich carbon atoms are in a linear or branched alkyl arrangement andwhere attachment is through the carbon of the keto group. Alkanoylgroups have the indicated number of carbon atoms, with the carbon of theketo group being included in the numbered carbon atoms. For example aC₂alkanoyl group is an acetyl group having the formula —(C═O)CH₃.Alkanoyl groups include, for example, C₂-C₈alkanoyl, C₂-C₆alkanoyl andC₂-C₄alkanoyl groups, which have from 2 to 8, from 2 to 6 or from 2 to 4carbon atoms, respectively. “C₁alkanoyl” refers to —(C═O)H, which (alongwith C₂-C₈alkanoyl) is encompassed by the term “C₁-C₈alkanoyl.”

An “alkanone” is a ketone group in which carbon atoms are in a linear orbranched alkyl arrangement. “C₃-C₈alkanone,” “C₃-C₆alkanone” and“C₃-C₄alkanone” refer to an alkanone having from 3 to 8, 6 or 4 carbonatoms, respectively. A C₃ alkanone has the structure —CH₂—(C═O)—CH₃.

Similarly, “alkyl ether” refers to a linear or branched ethersubstituent (i.e., an alkyl group that is substituted with an alkoxygroup). Alkyl ether groups include C₂-C₈alkyl ether, C₂-C₆alkyl etherand C₂-C₄alkyl ether groups, which have 2 to 8, 6 or 4 carbon atoms,respectively. A C₂ alkyl ether has the structure —CH₂—O—CH₃.

The term “alkoxycarbonyl” refers to an alkoxy group attached through aketo (—(C═O)—) bridge (i.e., a group having the general structure—C(═O)—O-alkyl). Alkoxycarbonyl groups include C₁-C₈, C₁-C₆ andC₁-C₄alkoxycarbonyl groups, which have from 1 to 8, 6 or 4 carbon atoms,respectively, in the alkyl portion of the group (i.e., the carbon of theketo bridge is not included in the indicated number of carbon atoms).“C₁alkoxycarbonyl” refers to —C(═O)—O—CH₃; C₃alkoxycarbonyl indicates—C(═O)—O—(CH₂)₂CH₃ or —C(═O)—O—(CH)(CH₃)₂.

“Alkanoyloxy,” as used herein, refers to an alkanoyl group linked via anoxygen bridge (i.e., a group having the general structure—O—C(═O)-alkyl). Alkanoyloxy groups include C₂-C₈, C₂-C₆ andC₂-C₄alkanoyloxy groups, which have from 2 to 8, 6 or 4 carbon atoms.“C₂alkanoyloxy” refers to —O—C(═O)—CH₃.

“Alkanoylamino,” as used herein, refers to an alkanoyl group attachedthrough an amino linker (i.e., a group having the general structure—N(R)—C(═O)-alkyl), in which R is hydrogen or C₁-C₆alkyl. Alkanoylaminogroups include C₂-C₈, C₂-C₆ and C₂-C₄alkanoylamino groups, which havefrom 2 to 8, 6 or 4 carbon atoms, respectively.

“Alkylsulfonyl” refers to groups of the formula —(SO₂)-alkyl, in whichthe sulfur atom is the point of attachment. Alkylsulfonyl groups includeC₁-C₆alkylsulfonyl and C₁-C₄alkylsulfonyl groups, which have from 1 to 6or from 1 to 4 carbon atoms, respectively. Methylsulfonyl is onerepresentative alkylsulfonyl group. “C₁-C₄haloalkylsulfonyl” is analkylsulfonyl group of from 1 to 4 carbon atoms that is substituted withat least one halogen (e.g., trifluoromethylsulfonyl).

“Alkylsulfonylamino” refers to groups of the formula —N(R)—(SO₂)-alkyl,in which R is hydrogen or C₁-C₆alkyl and the nitrogen atom is the pointof attachment. Alkylsulfonylamino groups include C₁-C₆alkylsulfonylaminoand C₁-C₄alkylsulfonylamino groups, which have from 1 to 6 or 1 to 4carbon atoms, respectively. Methylsulfonylamino is a representativealkylsulfonylamino group.

“Aminosulfonyl” refers to groups of the formula —(SO₂)—NH₂, in which thesulfur atom is the point of attachment. The term “mono- ordi-(C₁-C₆alkyl)aminosulfonyl” refers to groups that satisfy the formula—(SO₂)—NR₂, in which the sulfur atom is the point of attachment, and inwhich one R is C₁-C₈alkyl and the other R is hydrogen or anindependently chosen C₁-C₆alkyl.

“Alkylamino” refers to a secondary or tertiary amine of the formula—NH-alkyl or —N(alkyl)(alkyl), wherein each alkyl may be the same ordifferent. In this context, each alkyl may be linear, branched or cyclic(including (C₃-C₈cycloalkyl)C₀-C₄alkyl). Such groups include, forexample, mono- and di-(C₁-C₈alkyl)amino groups, in which each C₁-C₈alkylmay be the same or different, as well as mono- and di-(C₁-C₆alkyl)aminogroups and mono- and di-(C₁-C₄alkyl)amino groups.

“Alkylaminoalkyl” refers to an alkylamino group linked via an alkylenegroup (i.e., a group having the general structure -alkylene-NH-alkyl or-alkylene-N(alkyl)(alkyl)) in which each alkyl is selected independentlyfrom alkyl, cycloalkyl and (cycloalkyl)alkyl groups. Alkylaminoalkylgroups include, for example, mono- and di-(C₁-C₈alkyl)aminoC₁-C₈alkyl,mono- and di-(C₁-C₆alkyl)aminoC₁-C₆alkyl and mono- anddi-(C₁-C₆alkyl)aminoC₁-C₄alkyl. “Mono- ordi-(C₁-C₆alkyl)aminoC₀-C₈alkyl” refers to a mono- ordi-(C₁-C₆alkyl)amino group linked via a single covalent bond or aC₁-C₈alkylene group. The following are representative alkylaminoalkylgroups:

It will be apparent that the definition of “alkyl” as used in the terms“alkylamino” and “alkylaminoalkyl” differs from the definition of“alkyl” used for all other alkyl-containing groups, in the inclusion ofcycloalkyl and (cycloalkyl)alkyl groups (e.g.,(C₃-C₈cycloalkyl)C₀-C₄alkyl).

Similarly, “alkylaminoalkoxy” refers to an alkylamino group linked viaan alkoxy group (i.e., a group having the general structure—O-alkyl-NH-alkyl or —O-alkyl-N(alkyl)(alkyl)) in which each alkyl isselected independently. Such groups include, for example, mono- anddi-(C₁-C₆alkyl)aminoC₁-C₄alkoxy groups, such as

The term “aminocarbonyl” refers to an amide group (i.e., —(C═O)NH₂).“Mono- or di-(C₁-C₆alkyl)aminocarbonylC₀-C₄alkyl” is an aminocarbonylgroup in which one or both of the hydrogen atoms is replaced withC₁-C₆alkyl, and which is linked via a single covalent bond (i.e., mono-or di-(C₁-C₆alkyl)aminocarbonyl) or a C₁-C₄alkylene group (i.e.,—(C₀-C₄alkyl)-(C═O)N(C₁-C₈alkyl)₂). If both hydrogen atoms are soreplaced, the C₁-C₆alkyl groups may be the same or different.

The term “halogen” refers to fluorine, chlorine, bromine or iodine.

A “haloalkyl” is an alkyl group that is substituted with 1 or moreindependently chosen halogens (e.g., “C₁-C₈haloalkyl” groups have from 1to 8 carbon atoms; “C₁-C₆haloalkyl” groups have from 1 to 6 carbonatoms). Examples of haloalkyl groups include, but are not limited to,mono-, di- or tri-fluoromethyl; mono-, di- or tri-chloromethyl; mono-,di-, tri-, tetra- or penta-fluoroethyl; mono-, di-, tri-, tetra- orpenta-chloroethyl; and 1,2,2,2-tetrafluoro-1-trifluoromethyl-ethyl.Typical haloalkyl groups are trifluoromethyl and difluoromethyl. Theterm “haloalkoxy” refers to a haloalkyl group as defined above attachedvia an oxygen bridge. “C₁-C₈haloalkoxy” groups have 1 to 8 carbon atoms.

A dash (“-”) that is not between two letters or symbols is used toindicate a point of attachment for a substituent. For example, —CONH₂ isattached through the carbon atom.

A “carbocycle” or “carbocyclic group” comprises at least one ring formedentirely by carbon-carbon bonds (referred to herein as a carbocyclicring), and does not contain a heterocycle. Unless otherwise specified,each ring within a carbocycle may be independently saturated, partiallysaturated or aromatic, and is optionally substituted as indicated. Acarbocycle generally has from 1 to 3 fused, pendant or spiro rings;carbocycles within certain embodiments have one ring or two fused rings.Typically, each ring contains from 3 to 8 ring members (i.e., C₃-C₈);C₅-C₇ rings are recited in certain embodiments. Carbocycles comprisingfused, pendant or spiro rings typically contain from 9 to 14 ringmembers. Certain representative carbocycles are cycloalkyl as describedabove. Other carbocycles are aryl (i.e., contain at least one aromaticcarbocyclic ring, with or without one or more additional aromatic and/orcycloalkyl rings). Such aryl carbocycles include, for example, phenyl,naphthyl (e.g., 1-naphthyl and 2-naphthyl), fluorenyl, indanyl and1,2,3,4-tetrahydro-naphthyl.

Certain carbocycles recited herein are C₆-C₁₀arylC₀-C₈alkyl groups(i.e., groups in which a 6- to 10-membered carbocyclic group comprisingat least one aromatic ring is linked via a single covalent bond or aC₁-C₈alkylene group). Such groups include, for example, phenyl andindanyl, as well as groups in which either of the foregoing is linkedvia C₁-C₈alkylene, preferably via C₁-C₄alkylene. Phenyl groups linkedvia a single covalent bond or C₁-C₆alkylene group are designatedphenylC₀-C₆alkyl (e.g., benzyl, 1-phenyl-ethyl, 1-phenyl-propyl and2-phenyl-ethyl).

A “heterocycle” or “heterocyclic group” has from 1 to 3 fused, pendantor spiro rings, at least one of which is a heterocyclic ring (i.e., oneor more ring atoms is a heteroatom independently chosen from O, S and N,with the remaining ring atoms being carbon). Additional rings, ifpresent, may be heterocyclic or carbocyclic. Typically, a heterocyclicring comprises 1, 2, 3 or 4 heteroatoms; within certain embodiments eachheterocyclic ring has 1 or 2 heteroatoms per ring. Each heterocyclicring generally contains from 3 to 8 ring members (rings having from 4 or5 to 7 ring members are recited in certain embodiments) and heterocyclescomprising fused, pendant or spiro rings typically contain from 9 to 14ring members. Certain heterocycles comprise a sulfur atom as a ringmember; in certain embodiments, the sulfur atom is oxidized to SO orSO₂. Heterocycles may be optionally substituted with a variety ofsubstituents, as indicated. Unless otherwise specified, a heterocyclemay be a heterocycloalkyl group (i.e., each ring is saturated orpartially saturated) or a heteroaryl group (i.e., at least one ringwithin the group is aromatic), such as a 5- to 10-membered heteroaryl(which may be monocyclic or bicyclic) or a 6-membered heteroaryl (e.g.,pyridyl or pyrimidyl). N-linked heterocyclic groups are linked via acomponent nitrogen atom.

Heterocyclic groups include, for example, azepanyl, azocinyl,benzimidazolyl, benzimidazolinyl, benzisothiazolyl, benzisoxazolyl,benzofuranyl, benzothiofuranyl, benzoxazolyl, benzothiazolyl,benztetrazolyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl,dihydrofuro[2,3-b]tetrahydrofuranyl, dihydroisoquinolinyl,dihydrotetrahydrofuranyl, 1,4-dioxa-8-aza-spiro[4.5]decyl, dithiazinyl,furanyl, furazanyl, imidazolinyl, imidazolidinyl, imidazolyl, indazolyl,indolenyl, indolinyl, indolizinyl, indolyl, isobenzofuranyl,isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isothiazolyl,isoxazolyl, isoquinolinyl, morpholinyl, naphthyridinyl,octahydroisoquinolinyl, oxadiazolyl, oxazolidinyl, oxazolyl,phthalazinyl, piperazinyl, piperidinyl, piperidonyl, pteridinyl,purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl,pyridazinyl, pyridoimidazolyl, pyridooxazolyl, pyridothiazolyl, pyridyl,pyrimidyl, pyrrolidinyl, pyrrolidonyl, pyrrolinyl, pyrrolyl,quinazolinyl, quinolinyl, quinoxalinyl, quinuclidinyl,tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, thiadiazinyl,thiadiazolyl, thiazolyl, thienothiazolyl, thienooxazolyl,thienoimidazolyl, thienyl, thiophenyl, thiomorpholinyl and variantsthereof in which the sulfur atom is oxidized, triazinyl, and any of theforegoing that are substituted with from 1 to 4 substituents asdescribed above.

A “heterocycleC₀-C₈alkyl” is a heterocyclic group linked via a singlecovalent bond or C₁-C₈alkylene group. A (4- to 7-memberedheterocycle)C₀-C₈alkyl is a heterocyclic group (e.g., monocyclic orbicyclic) having from 4 to 7 ring members linked via a single covalentbond or an alkylene group having from 1 to 8 carbon atoms. A“(6-membered heteroaryl)C₀-C₆alkyl” refers to a heteroaryl group linkedvia a direct bond or C₁-C₆alkyl group.

Certain heterocycles are 4- to 12-membered, 5- to 10-membered, 3- to7-membered, 4- to 7-membered or 5- to 7-membered groups that contain 1heterocyclic ring or 2 fused, pendant or spiro rings, optionallysubstituted. 4- to 10-membered heterocycloalkyl groups include, forexample, piperidinyl, piperazinyl, pyrrolidinyl, azepanyl,1,4-dioxa-8-aza-spiro[4.5]dec-8-yl, morpholino, thiomorpholino and1,1-dioxo-thiomorpholin-4-yl. Such groups may be substituted asindicated. Representative aromatic heterocycles are azocinyl, pyridyl,pyrimidyl, imidazolyl, tetrazolyl and 3,4-dihydro-1H-isoquinolin-2-yl.

A “substituent,” as used herein, refers to a molecular moiety that iscovalently bonded to an atom within a molecule of interest. For example,a ring substituent may be a moiety such as a halogen, alkyl group,haloalkyl group or other group that is covalently bonded to an atom(preferably a carbon or nitrogen atom) that is a ring member.Substituents of aromatic groups are generally covalently bonded to aring carbon atom. The term “substitution” refers to replacing a hydrogenatom in a molecular structure with a substituent, such that the valenceon the designated atom is not exceeded, and such that a chemicallystable compound (i.e., a compound that can be isolated, characterized,and tested for biological activity) results from the substitution.

Groups that are “optionally substituted” are unsubstituted or aresubstituted by other than hydrogen at one or more available positions,typically 1, 2, 3, 4 or 5 positions, by one or more suitable groups(which may be the same or different). Optional substitution is alsoindicated by the phrase “substituted with from 0 to X substituents,”where X is the maximum number of possible substituents. Certainoptionally substituted groups are substituted with from 0 to 2, 3 or 4independently selected substituents (i.e., are unsubstituted orsubstituted with up to the recited maximum number of substitutents).

The terms “VR1” and “capsaicin receptor” are used interchangeably hereinto refer to a type 1 vanilloid receptor. Unless otherwise specified,these terms encompass both rat and human VR1 receptors (e.g., GenBankAccession Numbers AF327067, AJ277028 and NM_(—)018727; sequences ofcertain human VR1 cDNAs and the encoded amino acid sequences areprovided in U.S. Pat. No. 6,482,611), as well as homologues thereoffound in other species.

A “VR1 modulator,” also referred to herein as a “modulator,” is acompound that modulates VR1 activation and/or VR1-mediated signaltransduction. VR1 modulators specifically provided herein are compoundsof Formula I and pharmaceutically acceptable salts thereof. Certainpreferred VR1 modulators are not vanilloids. A VR1 modulator may be aVR1 agonist or antagonist. A modulator binds with high affinity if theK_(i) at VR1 is less than 1 micromolar, preferably less than 500nanomolar, 100 nanomolar, 10 nanomolar or 1 nanomolar. A representativeassay for determining K_(i) at VR1 is provided in Example 5, herein.

A modulator is considered an “antagonist” if it detectably inhibitsvanilloid ligand binding to VR1 and/or VR1-mediated signal transduction(using, for example, the representative assay provided in Example 6); ingeneral, such an antagonist inhibits VR1 activation with a IC₅₀ value ofless than 1 micromolar, preferably less than 500 nanomolar, and morepreferably less than 100 nanomolar, 10 nanomolar or 1 nanomolar withinthe assay provided in Example 6. VR1 antagonists include neutralantagonists and inverse agonists.

A “neutral antagonist” of VR1 is a compound that inhibits the activityof vanilloid ligand at VR1, but does not significantly change the basalactivity of the receptor (i.e., within a calcium mobilization assay asdescribed in Example 6 performed in the absence of vanilloid ligand, VR1activity is reduced by no more than 10%, more preferably by no more than5%, and even more preferably by no more than 2%; most preferably, thereis no detectable reduction in activity). Neutral antagonists of VR1 mayalso inhibit the binding of vanilloid ligand to VR1.

An “inverse agonist” of VR1 is a compound that reduces the activity ofVR1 below its basal activity level in the absence of added vanilloidligand. Inverse agonists of VR1 may also inhibit the activity ofvanilloid ligand at VR1 and/or binding of vanilloid ligand to VR1. Thebasal activity of VR1, as well as the reduction in VR1 activity due tothe presence of VR1 antagonist, may be determined from a calciummobilization assay, as described in Example 6.

As used herein a “capsaicin receptor agonist” or “VR1 agonist” is acompound that elevates the activity of the receptor above the basalactivity level (i.e., enhances VR1 activation and/or VR1-mediated signaltransduction). Capsaicin receptor agonist activity may be identifiedusing the representative assay provided in Example 6. In general, suchan agonist has an EC₅₀ value of less than 1 micromolar, preferably lessthan 500 nanomolar, and more preferably less than 100 nanomolar or 10nanomolar within the assay provided in Example 6.

A “vanilloid” is capsaicin or any capsaicin analogue that comprises aphenyl ring with two oxygen atoms bound to adjacent ring carbon atoms(one of which carbon atom is located para to the point of attachment ofa third moiety that is bound to the phenyl ring). A vanilloid is a“vanilloid ligand” if it binds to VR1 with a K_(i) (determined asdescribed herein) that is no greater than 10 μM. Vanilloid ligandagonists include capsaicin, olvanil, N-arachidonoyl-dopamine andresiniferatoxin (RTX). Vanilloid ligand antagonists include capsazepineand iodo-resiniferatoxin.

A “therapeutically effective amount” (or dose) is an amount that, uponadministration to a patient, results in a discernible patient benefit(e.g., provides detectable relief from a condition being treated). Suchrelief may be detected using any appropriate criteria, includingalleviation of one or more symptoms such as pain. A therapeuticallyeffective amount or dose generally results in a concentration ofcompound in a body fluid (such as blood, plasma, serum, CSF, synovialfluid, lymph, cellular interstitial fluid, tears or urine) that issufficient to alter the binding of vanilloid ligand to VR1 in vitro(using the assay provided in Example 5) and/or VR1-mediated signaltransduction (using an assay provided in Example 6). It will be apparentthat the discernible patient benefit may be apparent afteradministration of a single dose, or may become apparent followingrepeated administration of the therapeutically effective dose accordingto a predetermined regimen, depending upon the indication for which thecompound is administered.

A “patient” is any individual treated with a compound provided herein.Patients include humans, as well as other animals such as companionanimals (e.g., dogs and cats) and livestock. Patients may beexperiencing one or more symptoms of a condition responsive to capsaicinreceptor modulation (e.g., pain, exposure to vanilloid ligand, itch,urinary incontinence, overactive bladder, respiratory disorders, coughand/or hiccup), or may be free of such symptom(s) (i.e., treatment maybe prophylactic in a patient considered at risk for the development ofsuch symptoms).

Substituted Biaryl Piperazinyl-Pyridine Analogues

As noted above, the present invention provides substituted biarylpiperazinyl-pyridine analogues that may be used in a variety ofcontexts, including in the treatment of pain (e.g., neuropathic orperipheral nerve-mediated pain); exposure to capsaicin; exposure toacid, heat, light, tear gas, air pollutants (such as, for example,tobacco smoke), infectious agents (including viruses, bacteria andyeast), pepper spray or related agents; respiratory conditions such asasthma or chronic obstructive pulmonary disease; itch; urinaryincontinence or overactive bladder; cough or hiccup; and/or obesity.Such compounds may also be used within in vitro assays (e.g., assays forreceptor activity), as probes for detection and localization of VR1 andas standards in ligand binding and VR1-mediated signal transductionassays.

Certain compounds provided herein detectably modulate the binding ofcapsaicin to VR1 at nanomolar (i.e., submicromolar) concentrations, atsubnanomolar concentrations, or at concentrations below 100 picomolar,20 picomolar, 10 picomolar or 5 picomolar. Such modulators arepreferably not vanilloids. Certain modulators are VR1 antagonists andhave no detectable agonist activity in the assay described in Example 6.Preferred VR1 modulators further bind with high affinity to VR1.

In certain embodiments, substituted biaryl piperazinyl-pyridineanalogues further satisfy Formula II:

wherein:

-   D, K, J and F are independently N, CH or carbon substituted with a    substituent represented by R₁ or R₁₀;-   R₁ represents from 0 to 3 substituents independently chosen from:    -   (a) halogen, cyano and nitro;    -   (b) groups of the formula -Q-M-R_(y); and    -   (c) groups that are taken together with R₁₀ to form a fused 5-        to 7-membered carbocyclic or heterocyclic ring that is        substituted with from 0 to 4 substituents independently chosen        from halogen, cyano, nitro and groups of the formula -Q-M-R_(y);-   R₁₀ represents one substituent chosen from:    -   (a) groups of the formula -Q-M-R_(y); and    -   (b) groups that are taken together with a R₁ to form a fused,        optionally substituted 5- to 7-membered carbocyclic or        heterocyclic ring;    -   such that R₁₀ is not hydroxy, amino or an unsubstituted group        chosen from C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆alkoxy,        C₂-C₆alkyl ether, C₂-C₆alkanoyl, C₃-C₆alkanone, C₁-C₆haloalkyl,        C₁-C₆haloalkoxy, mono- or di-(C₁-C₆alkyl)amino,        C₁-C₆alkylsulfonyl, mono- or di-(C₁-C₆alkyl)aminosulfonyl or        mono- or di-(C₁-C₆alkyl)aminocarbonyl;        and the remaining variables are as described above for Formula        I.

In certain embodiments, substituted biaryl piperazinyl-pyridineanalogues of Formula I satisfy one or more of Formulas IIa-IIf, in whichvariables are as indicated for Formula II, except as defined below:

Within Formula IIe and Formula IIf:

-   R_(1a) and R_(1b) are independently hydrogen, halogen, amino, cyano,    C₁-C₆alkyl, C₁-C₆haloalkyl, C₁-C₆alkoxy, C₁-C₆alkylsulfonyl, mono-    or di-(C₁-C₆alkyl)aminosulfonyl, (C₃-C₈cycloalkyl)aminocarbonyl or a    group of the formula:

-    wherein

-    represents a 4- to 7-membered, N-linked heterocycloalkyl; and-   R₁₀ is:    -   (a) —COOH, aminocarbonyl, imino, C₁-C₆alkoxycarbonyl or        C₂-C₆alkanoyl;    -   (b) C₁-C₆alkyl, C₂-C₆alkyl ether, C₁-C₆alkoxy, C₁-C₆alkylamino        or mono- or di-(C₁-C₄alkyl)aminocarbonyl, each of which is        substituted with from 1 to 4 substituents independently chosen        from:        -   (i) halogen, hydroxy, —COOH, cyano, amino and aminocarbonyl;            and        -   (ii) C₁-C₆alkoxy, C₃-C₈cycloalkyl, C₁-C₆alkylsulfonyl,            C₁-C₆alkoxycarbonyl, C₂-C₄alkanoyl, mono- and            di-(C₁-C₈alkyl)amino, phenyl and 4- to 7-membered            heterocycles, each of which is substituted with from 0 to 4            substituents independently chosen from halogen, C₁-C₄alkyl            and C₁-C₆alkoxy; or    -   (c) (C₃-C₈cycloalkyl)aminocarbonyl or a group of the formula:

-   -    wherein

-   -    represents a 4- to 7-membered, N-linked heterocycloalkyl, each        of which is substituted with from 0 to 4 substituents        independently chosen from hydroxy, —COOH, cyano, amino,        aminocarbonyl, C₁-C₆alkyl, C₂-C₆alkyl ether, C₃-C₈cycloalkyl,        C₁-C₆alkoxy, C₁-C₆alkylsulfonyl, C₁-C₆alkoxycarbonyl,        C₂-C₄alkanoyl, C₂-C₄alkanoylamino and mono- and        di-(C₁-C₄alkyl)amino.

Within certain embodiments of Formulas IIa-IIf, Ar₂ is phenyl, pyridylor pyrimidyl, each of which is substituted with from 1 to 3 substituentsindependently chosen from amino, cyano, halogen, C₁-C₄alkyl,C₁-C₄haloalkyl, C₁-C₄-aminoalkyl, C₁-C₄alkoxy, C₁-C₄haloalkoxy,C₁-C₄alkylthio, and mono- and di-(C₁-C₄alkyl)aminoC₀-C₄alkyl; and R₄represents 0 substituents or one methyl substituent.

In further embodiments, substituted biaryl piperazinyl-pyridineanalogues of Formula I further satisfy Formula III:

wherein R₁, K, J and F are as described for Formula II and the remainingvariables are as described above for Formula I.

In certain embodiments, substituted biaryl piperazinyl-pyridineanalogues of Formula III further satisfy one or more of FormulasIIIa-IIId, in which variables are as indicated for Formula III:

Certain substituted biaryl piperazinyl-pyridine analogues of Formula IIIfurther satisfy Formula IIIe or IIIf:

wherein:

-   A, B, E and T are independently nitrogen or CR_(2a);-   R_(1a) and R_(1b) are independently hydrogen, halogen, amino, cyano,    C₁-C₆alkyl, C₁-C₆haloalkyl, C₁-C₆alkoxy, C₁-C₆alkylsulfonyl, mono-    or di-(C₁-C₆alkyl)aminosulfonyl, (C₃-C₈cycloalkyl)aminocarbonyl or a    group of the formula:

-    wherein

-    represents a 4- to 7-membered, N-linked heterocycloalkyl;-   Each R_(2a) is independently chosen from hydrogen, hydroxy, amino,    cyano, halogen, aminosulfonyl, aminocarbonyl, —COOH, C₁-C₄alkyl,    C₁-C₄haloalkyl, C₁-C₄hydroxyalkyl, C₁-C₄alkoxy, C₁-C₄alkylthio,    C₁-C₄haloalkoxy, mono- and di-(C₁-C₈alkyl)aminoC₀-C₄alkyl,    C₂-C₄alkanoyl, C₁-C₄alkoxycarbonyl, C₁-C₄alkylsulfonyl,    C₁-C₄haloalkylsulfonyl and mono- or di-(C₁-C₄alkyl)aminosulfonyl,    such that at least one R_(2a) is not hydrogen;-   R₄ is hydrogen, methyl, ethyl or oxo;-   R₁₀ is:    -   (a) nitro, —COOH, aminocarbonyl, imino, C₁-C₆alkoxycarbonyl or        C₂-C₆alkanoyl;    -   (b) C₁-C₆alkyl, C₂-C₆alkyl ether, C₁-C₆alkoxy, C₁-C₆alkylamino        or mono- or di-(C₁-C₄alkyl)aminocarbonyl, each of which is        substituted with from 1 to 4 substituents independently chosen        from:        -   (i) halogen, hydroxy, —COOH, cyano, amino and aminocarbonyl;            and        -   (ii) C₁-C₆alkoxy, C₃-C₈cycloalkyl, C₁-C₆alkylsulfonyl,            C₁-C₆alkoxycarbonyl, C₂-C₄alkanoyl, mono- and            di-(C₁-C₈alkyl)amino, phenyl and 4- to 7-membered            heterocycles, each of which is substituted with from 0 to 4            substituents independently chosen from halogen, C₁-C₄alkyl            and C₁-C₆alkoxy; or    -   (c) (C₃-C₈cycloalkyl)aminocarbonyl or a group of the formula:

-   -    wherein

-   -    represents a 4- to 7-membered, N-linked heterocycloalkyl, each        of which is substituted with from 0 to 4 substituents        independently chosen from hydroxy, —COOH, cyano, amino,        aminocarbonyl, C₁-C₆alkyl, C₂-C₆alkyl ether, C₃-C₈cycloalkyl,        C₁-C₆alkoxy, C₁-C₆alkylsulfonyl, C₁-C₆alkoxycarbonyl,        C₂-C₄alkanoyl, C₂-C₄alkanoylamino and mono- and        di-(C₁-C₄alkyl)amino;

-   R_(x) is independently selected at each occurrence from hydrogen,    methyl, amino and cyano;    and the remaining variables are as described for Formula III.

Certain substituted biaryl piperazinyl-pyridine analogues of FormulaIIIe further satisfy Formula IIIg, in which R₄ is hydrogen or methyl andthe remaining variables are as described for Formula IIIe:

Within certain embodiments of Formulas IIIe, IIIf and IIIg:

-   R_(1a) is halogen, cyano, methyl or trifluoromethyl;-   R_(1b) is hydrogen, halogen, amino, C₁-C₃alkyl, C₁-C₃alkoxy, mono-    or di-(C₁-C₄alkyl)amino, mono- or di-(C₁-C₄alkyl)aminocarbonyl,    C₁-C₃alkylsulfonyl, or mono- or di-(C₁-C₄alkyl)aminosulfonyl; and-   each R_(2a) is independently chosen from hydrogen, halogen, cyano,    amino, C₁-C₄alkyl, C₁-C₄alkoxy, and C₁-C₄haloalkyl.    -   Within certain embodiments of Formulas IIIe and IIIg, R₅ and R₆        are independently chosen from:    -   (i) hydrogen; and    -   (ii) C₁-C₆alkyl, C₂-C₆alkenyl, (C₅-C₇cycloalkyl)C₀-C₂alkyl and        groups that are joined to L to form a 4- to 7-membered        heterocycle; each of which is substituted with from 0 to 2        substituents independently chosen from halogen, hydroxy, oxo,        —COOH, aminocarbonyl, aminosulfonyl, C₁-C₄alkyl, C₁-C₄haloalkyl,        C₁-C₄alkoxy, C₁-C₄alkoxycarbonyl, C₁-C₄alkylsulfonyl and mono-        and di-(C₁-C₆alkyl)amino;-   or R₅ and R₆, together with the N to which they are bound, form a 4-    to 7-membered heterocycloalkyl that is substituted with from 0 to 2    substituents independently chosen from halogen, hydroxy, amino, oxo,    aminocarbonyl, aminosulfonyl, —COOH, C₁-C₄alkyl, C₁-C₄alkoxy,    C₁-C₄hydroxyalkyl, C₂-C₄alkyl ether, C₁-C₄alkoxycarbonyl,    C₂-C₄alkanoyl, C₂-C₄alkanoylamino, C₁-C₄alkylsulfonyl, and mono- and    di-(C₁-C₄alkyl)aminoC₀-C₂alkyl.

In further embodiments, substituted biaryl piperazinyl-pyridineanalogues of Formula I further satisfy Formula IV:

wherein:

-   F is carbon substituted with a substituent represented by R₁ or R₁₀;-   R₁₀ is as described for Formula III;    and the remaining variables are as described for Formula II.

Certain substituted biaryl piperazinyl-pyridine analogues of Formula IVfurther satisfy Formula IVa, in which variables are as described forFormula IV:

Certain substituted biaryl piperazinyl-pyridine analogues of Formula IVfurther satisfy Formula IVb or Formula IVc:

wherein:

-   A, B, E and T are independently nitrogen or CR_(2a);-   R_(1a) is halogen, amino, cyano, C₁-C₆alkyl, C₁-C₆haloalkyl,    C₁-C₆alkoxy, C₁-C₆alkylsulfonyl or mono- or    di-(C₁-C₆alkyl)aminosulfonyl;-   R_(1b) is hydrogen, halogen, amino, cyano, C₁-C₆alkyl,    C₁-C₆haloalkyl, C₁-C₆alkoxy, C₁-C₆alkylsulfonyl, mono- or    di-(C₁-C₆alkyl)aminosulfonyl, (C₃-C₈cycloalkyl)aminocarbonyl or a    group of the formula:

-    wherein

-    represents a 4- to 7-membered, N-linked heterocycloalkyl;-   Each R_(2a) is independently chosen from hydrogen, hydroxy, amino,    cyano, halogen, aminosulfonyl, aminocarbonyl, —COOH, C₁-C₄alkyl,    C₁-C₄haloalkyl, C₁-C₄hydroxyalkyl, C₁-C₄alkoxy, C₁-C₄alkylthio,    C₁-C₄haloalkoxy, mono- and di-(C₁-C₈alkyl)aminoC₀-C₄alkyl,    C₂-C₄alkanoyl, C₁-C₄alkoxycarbonyl, C₁-C₄alkylsulfonyl,    C₁-C₄haloalkylsulfonyl and mono- and di-(C₁-C₄alkyl)aminosulfonyl,    such that at least one R_(2a) is not hydrogen;-   R₄ is hydrogen, methyl, ethyl or oxo;-   R₁₀ is as described for Formula IIIe and Formula IIIf;-   R_(x) is independently selected at each occurrence from hydrogen,    methyl, amino and cyano; and the remaining variables are as    described for Formula IV.

Within certain embodiments of Formula IVa and Formula IVb:

-   R_(1a) is halogen, cyano, methyl or trifluoromethyl;-   R_(1b) is hydrogen, halogen, amino, C₁-C₃alkyl, C₁-C₃alkoxy, mono-    or di-(C₁-C₄alkyl)amino, mono- or di-(C₁-C₄alkyl)aminocarbonyl,    C₁-C₃alkylsulfonyl, or mono- or di-(C₁-C₄alkyl)aminosulfonyl;-   each R_(2a) is independently chosen from hydrogen, halogen, cyano,    amino, C₁-C₄alkyl, C₁-C₄alkoxy, and C₁-C₄haloalkyl; and-   R₅ and R₆ are independently chosen from:    -   (i) hydrogen; and    -   (ii) C₁-C₆alkyl, C₂-C₆alkenyl, (C₅-C₇cycloalkyl)C₀-C₂alkyl and        groups that are joined to L to form a 4- to 7-membered        heterocycle; each of which is substituted with from 0 to 2        substituents independently chosen from halogen, hydroxy, oxo,        —COOH, aminocarbonyl, aminosulfonyl, C₁-C₄alkyl, C₁-C₄haloalkyl,        C₁-C₄alkoxy, C₁-C₄alkoxycarbonyl, C₁-C₄alkylsulfonyl and mono-        and di-(C₁-C₆alkyl)amino;-   or R₅ and R₆, together with the N to which they are bound, form a 4-    to 7-membered heterocycloalkyl that is substituted with from 0 to 2    substituents independently chosen from halogen, hydroxy, amino, oxo,    aminocarbonyl, aminosulfonyl, —COOH, C₁-C₄alkyl, C₁-C₄alkoxy,    C₁-C₄hydroxyalkyl, C₂-C₄alkyl ether, C₁-C₄alkoxycarbonyl,    C₂-C₄alkanoyl, C₂-C₄alkanoylamino, C₁-C₄alkylsulfonyl, and mono- and    di-(C₁-C₄alkyl)aminoC₀-C₂alkyl.

Further substituted biaryl piperazinyl-pyridine analogues of Formula Iadditionally satisfy one of Formulas V, VI, VII or VIII:

wherein:

-   D, K, G, J and F are independently N, CH or carbon substituted with    a substituent represented by R₁ or R₁₀;-   R₁ is as described for Formula II;-   R₁₀ is as described for Formula III;    and the remaining variables are as described for Formula I.    Within certain embodiments of the above Formulas, variables are as    follows:-   Ar₁, R₁, R_(1a), R_(1b), R_(1c) and R₁₀    -   Certain Ar₁ groups satisfy the formula:

-   -    in which variables are as described above. In certain        embodiments, D is N; in further embodiments, J, F, K and/or G        are N; within still further embodiments, D and F are both N.        Within other embodiments, K is N or both K and J are N. In still        further embodiments, J, F, K and/or G are optionally substituted        carbon (e.g., J, F and K are optionally substituted carbon).        Within certain compounds, F is substituted carbon (i.e., one        substituent represented by R₁ or R₁₀ is located ortho to the        point of attachment) and/or one substituent represented by R₁ or        R₁₀ is located meta or para to the point of attachment, wherein        the point of attachment refers to the attachment to the        piperazinyl ring.

Certain substituents satisfy the formula Q-M-Ry. It will be apparentthat if Q is C₀ and M is a single covalent bond, then R_(y) is directlylinked (via a single covalent bond) to the Ar₁ core ring.

R₁₀ in the above Formulas represents one substituent of Ar₁. In otherwords, a R₁₀ moiety, as defined above, is covalently bonded to any ringcarbon atom of Ar₁, such as a carbon atom at the position designated D,K, G, J or F in certain formulas. Within certain compounds of Formula IIand subformulas thereof, R₁₀ represents:

-   (a) —COOH, aminocarbonyl, imino, C₁-C₆alkoxycarbonyl or    C₂-C₆alkanoyl;-   (b) C₁-C₆alkyl, C₂-C₆alkyl ether, C₁-C₆alkoxy, C₁-C₆alkylamino or    mono- or di-(C₁-C₄alkyl)aminocarbonyl, each of which is substituted    with from 1 to 4 substituents independently chosen from:    -   (i) halogen, hydroxy, —COOH, cyano, amino and aminocarbonyl; and    -   (ii) C₁-C₆alkoxy, C₃-C₈cycloalkyl, C₁-C₆alkylsulfonyl,        C₁-C₆alkoxycarbonyl, C₂-C₄alkanoyl, mono- and        di-(C₁-C₈alkyl)amino, 4- to 7-membered heterocycles and phenyl,        each of which is substituted with from 0 to 4 substituents        independently chosen from halogen, C₁-C₄alkyl and C₁-C₆alkoxy;        or-   (c) (C₃-C₈cycloalkyl)aminocarbonyl or a group of the formula:

-    wherein

-    represents a 4- to 7-membered, N-linked heterocycloalkyl, each of    which is substituted with from 0 to 4 substituents independently    chosen from hydroxy, —COOH, cyano, amino, aminocarbonyl, C₁-C₆alkyl,    C₂-C₆alkyl ether, C₃-C₈cycloalkyl, C₁-C₆alkoxy, C₁-C₆alkylsulfonyl,    C₁-C₆alkoxycarbonyl, C₂-C₄alkanoyl, C₂-C₄alkanoylamino and mono- and    di-(C₁-C₄alkyl)amino.    Within certain compounds of the other Formulas, R₁₀ represents:-   (a) nitro, —COOH, aminocarbonyl, imino, C₁-C₆alkoxycarbonyl or    C₂-C₆alkanoyl;-   (b) C₁-C₆alkyl, C₂-C₆alkyl ether, C₁-C₆alkoxy, C₁-C₆alkylamino or    mono- or di-(C₁-C₄alkyl)aminocarbonyl, each of which is substituted    with from 1 to 4 substituents independently chosen from:    -   (i) halogen, hydroxy, —COOH, cyano, amino and aminocarbonyl; and    -   (ii) C₁-C₆alkoxy, C₃-C₈cycloalkyl, C₁-C₆alkylsulfonyl,        C₁-C₆alkoxycarbonyl, C₂-C₄alkanoyl, mono- and        di-(C₁-C₈alkyl)amino, 4- to 7-membered heterocycles and phenyl,        each of which is substituted with from 0 to 4 substituents        independently chosen from halogen, C₁-C₄alkyl and C₁-C₆alkoxy;        or-   (c) (C₃-C₈cycloalkyl)aminocarbonyl or a group of the formula:

-    wherein

-    represents a 4- to 7-membered, N-linked heterocycloalkyl, each of    which is substituted with from 0 to 4 substituents independently    chosen from hydroxy, —COOH, cyano, amino, aminocarbonyl, C₁-C₆alkyl,    C₂-C₆alkyl ether, C₃-C₈cycloalkyl, C₁-C₆alkoxy, C₁-C₆alkylsulfonyl,    C₁-C₆alkoxycarbonyl, C₂-C₄alkanoyl, C₂-C₄alkanoylamino and mono- and    di-(C₁-C₄alkyl)amino.    Representative R₁₀ groups include, for example, nitro,    aminocarbonyl, —COOH, C₁-C₄alkoxycarbonyl, hydroxyC₁-C₄alkyl (e.g.,    hydroxymethyl), aminoC₁-C₄alkyl (e.g., aminomethyl), cyanoC₁-C₄alkyl    (e.g., cyanomethyl), carboxyC₁-C₄alkyl (e.g., carboxymethyl), mono-    and di-(C₁-C₄alkyl)aminoC₁-C₄alkyl and mono- and    di-(C₁-C₄alkyl)aminoC₁-C₄alkoxy. Other representative R₁₀ groups    include (C₃-C₈cycloalkyl)aminocarbonyl groups and mono- and    di-(C₁-C₄alkyl)aminocarbonyl groups that are substituted with    C₁-C₄alkoxy, C₁-C₄haloalkyl C₂-C₆alkyl ether, mono- or    di-(C₁-C₄alkyl)amino, optionally substituted 5- to 7-membered    heterocycloalkyl groups (e.g., tetrahydrofuranyl or thiophenyl),    and/or optionally substituted phenyl. Additional representative R₁₀    groups include groups of the formula:

-    wherein

-    represents optionally substituted azetidinyl, pyrrolidinyl,    piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl or azepanyl.

R₁ in the above Formulas represents up to three optional substituents ofAr₁ (in addition to R₁₀). In certain embodiments, R₁ represents onesubstituent (i.e., Ar₁ is di-substituted). Representative R₁ groupsinclude, for example, halogen, nitro, cyano, methyl,C₁-C₄alkoxycarbonyl, trifluoromethyl and methylsulfonyl.

The variables R_(1a) and R_(1b), within Formulas that recite suchvariables, are generally as described above. Within certain compounds,R_(1a) and R_(1b) are independently hydrogen, halogen, amino, cyano,C₁-C₆alkyl, C₁-C₆haloalkyl, C₁-C₆alkoxy, C₁-C₆alkylsulfonyl, mono- ordi-(C₁-C₆alkyl)aminosulfonyl, (C₃-C₈cycloalkyl)aminocarbonyl or a groupof the formula:

-    wherein

-    represents a 4- to 7-membered, N-linked heterocycloalkyl. In    further embodiments, R_(1a) is nitro, —COOH, halogen, cyano, methyl    or trifluoromethyl; in further embodiments, R_(1a) is halogen,    cyano, methyl or trifluoromethyl.-   Ar₂, R₂ and R_(2a)

In certain substituted biaryl piperazinyl-pyridine analogues of theabove Formulas, Ar₂ is optionally substituted phenyl, pyridyl (i.e.,2-pyridyl, 3-pyridyl or 4-pyridyl) or pyrimidyl. In other compounds, Ar₂is a 9- to 12-membered bicyclic aryl or heteroaryl group that isoptionally substituted as described above. In some embodiments, Ar₂ isunsubstituted phenyl or unsubstituted pyridyl. In other embodiments, Ar₂is substituted with from 0 to 3 or from 1 to 3 substituentsindependently chosen from R₂ as described above. In certain suchcompounds, Ar₂ has at least one substituent (R₂) and each R₂ isindependently chosen from amino, cyano, halogen, aminosulfonyl,C₁-C₄alkyl, C₁-C₄haloalkyl, C₁-C₄hydroxyalkyl, C₁-C₄-aminoalkyl,C₁-C₄alkoxy, C₁-C₄alkylthio, C₁-C₄haloalkoxy, mono- anddi-(C₁-C₄alkyl)aminoC₀-C₄alkyl, C₂-C₄alkanoyl, C₁-C₄alkoxycarbonyl,C₁-C₄alkylsulfonyl, C₁-C₄haloalkylsulfonyl and mono- anddi-(C₁-C₄alkyl)aminosulfonyl. Representative substituents of Ar₂ includeamino, cyano, halogen, C₁-C₄alkyl, C₁-C₄haloalkyl, C₁-C₄-aminoalkyl,C₁-C₄alkoxy, C₁-C₄haloalkoxy, C₁-C₄alkylthio and mono- anddi-(C₁-C₄alkyl)aminoC₀-C₄alkyl. In certain such compounds, Ar₂ issubstituted meta and/or para to the point of attachment, wherein thepoint of attachment refers to the attachment to the core ring. In otherwords, if Ar₂ is phenyl, the phenyl is mono-substituted at the3-position, mono-substituted at the 4-position, or di-substituted andthe 3- and 4-positionsin such compounds. Representative Ar₂ groupsinclude phenyl, pyridyl and pyrimidyl, each of which is substituted withfrom 0 to 3 or from 1 to 3 substituents as described herein.

Within certain Ar₂ groups, one R₂ is taken together with an adjacent R₂to form a fused carbocycle or heterocycle. Representative such groupsinclude, for example, the following bicyclic groups, optionallysubstituted as described herein:

as well as variants of the foregoing in which the fused ring containsone or more additional double bonds, such as:

The variable R_(2a), when present, is generally as described above; incertain embodiments, each R_(2a) is independently chosen from hydrogen,hydroxyl, amino, cyano, halogen, aminosulfonyl, aminocarbonyl, —COOH,C₁-C₄alkyl, C₁-C₄haloalkyl, C₁-C₄hydroxyalkyl, C₁-C₄alkoxy,C₁-C₄alkylthio, C₁-C₄haloalkoxy, mono- anddi-(C₁-C₄alkyl)aminoC₀-C₄alkyl, C₂-C₄alkanoyl, C₁-C₄alkoxycarbonyl,C₁-C₄alkylsulfonyl, C₁-C₄haloalkylsulfonyl and mono- anddi-(C₁-C₄alkyl)aminosulfonyl. In further such compounds, each R_(2a) isindependently chosen from hydrogen, halogen, cyano, amino, C₁-C₄alkyl,C₁-C₄haloalkyl and C₁-C₄alkoxy. In still further such compounds, A is CHor CR_(2a), and each R_(2a) is independently chosen from cyano, halogen,C₁-C₄alkyl, C₁-C₄haloalkyl, C₁-C₄alkoxy and mono- anddi-(C₁-C₄alkyl)aminoC₀-C₂alkyl. In other such compounds, at least one ofA, B, E and T is N. Certain Ar₂ groups have the formula:

wherein B and E and R₂ are as described above.

-   R₃

In the definition of R₃, the variable “L” is defined as C₀-C₆alkylene orC₁-C₆alkylene that is taken together with R₅, R₆ or R₇ to form a 4- to7-membered heterocycle. In any heterocycle so formed, at least onecarbon atom present in L is also a ring atom, and is covalently bondedto a component atom of R₅, R₆ or R₇. The resulting heterocycle may be aheterocycloalkyl group (e.g., tetrahydrofuranyl, morpholinyl,piperidinyl or piperazinyl) or a heteroaryl group, such as pyridyl orpyrimidyl. R₃ groups comprising such a heterocycle include, for example:

R₃, in certain embodiments of various Formulas provided herein, is agroup of the formula:

wherein:

-   L is C₀-C₃alkylene; and-   R₅ and R₆ are:    -   (a) independently chosen from hydrogen, C₁-C₆alkyl,        C₂-C₆alkenyl, (C₃-C₈cycloalkyl)C₀-C₄alkyl, C₂-C₄alkanoyl and        groups that are joined to L to form a 4- to 7-membered        heterocycle; or    -   (b) joined to form a 4- to 12-membered heterocycloalkyl;    -   wherein each alkyl, alkenyl, (cycloalkyl)alkyl, alkanoyl and        heterocycloalkyl is substituted with from 0 to 4 substituents        independently chosen from (i) halogen, hydroxy, amino,        aminocarbonyl, oxo, —COOH and aminosulfonyl; and (ii)        C₁-C₄alkyl, C₅-C₇cycloalkyl, C₁-C₄alkoxy, C₂-C₄alkanoyl,        C₁-C₄haloalkyl, mono- and di-(C₁-C₄alkyl)aminoC₀-C₂alkyl, mono-        and di-(C₁-C₄alkyl)aminocarbonylC₀-C₂alkyl, phenylC₀-C₄alkyl and        (4- to 7-membered heterocycle)C₀-C₂alkyl, each of which is        substituted with from 0 to 4 secondary substituents        independently chosen from halogen, hydroxy, cyano, C₁-C₄alkyl,        C₁-C₄alkoxy and C₁-C₄haloalkyl.

Within certain such compounds, R₅ and R₆ are independently chosen from:

-   -   (i) hydrogen; and    -   (ii) C₁-C₆alkyl, C₂-C₆alkenyl, (C₅-C₇cycloalkyl)C₀-C₂alkyl and        groups that are joined to L to form a 4- to 7-membered        heterocycle; each of which is substituted with from 0 to 2        substituents independently chosen from halogen, hydroxy, oxo,        —COOH, aminocarbonyl, aminosulfonyl, C₁-C₄alkyl, C₁-C₄haloalkyl,        C₁-C₄alkoxy, C₁-C₄alkoxycarbonyl, C₁-C₄alkylsulfonyl and mono-        and di-(C₁-C₆alkyl)amino;    -   or R₅ and R₆, together with the N to which they are bound, form        a 4- to 7-membered heterocycloalkyl that is substituted with        from 0 to 2 substituents independently chosen from halogen,        hydroxy, amino, oxo, aminocarbonyl, aminosulfonyl, —COOH,        C₁-C₄alkyl, C₁-C₄alkoxy, C₁-C₄hydroxyalkyl, C₂-C₄alkyl ether,        C₁-C₄alkoxycarbonyl, C₂-C₄alkanoyl, C₂-C₄alkanoylamino,        C₁-C₄alkylsulfonyl, and mono- and di-(C₁-C₄alkyl)aminoC₀-C₂alkyl

Such R₃ groups include, for example, mono- and di-(C₁-C₄alkyl)aminogroups that are substituted with from 0 to 4 substituents independentlychosen from halogen, hydroxy, amino, oxo, aminocarbonyl, —COOH,aminosulfonyl, C₁-C₄alkyl, C₂-C₄alkenyl, C₅-C₇cycloalkyl,C₁-C₄haloalkyl, C₁-C₄alkoxy, C₂-C₄alkyl ether, C₂-C₄alkanoyl,C₁-C₄alkylsulfonyl, C₂-C₄alkanoylamino and mono- anddi-(C₁-C₄alkyl)amino. Representative such groups include:

Within other R₃ groups, R₅ and R₆, together with the N to which they arebound, form a pyrrolidine, piperazine, piperidine, azetidine ormorpholine ring, each of which is substituted with from 0 to 2substituents independently chosen from C₁-C₄alkyl and C₁-C₄hydroxyalkyl.

Still further R₃ groups include phenyl and 4- to 7-memberedheterocycles, each of which is substituted with from 0 to 4 substituentsindependently chosen from (a) halogen, hydroxy, amino, oxo,aminocarbonyl, aminosulfonyl and —COOH; and (b) C₁-C₄alkyl,C₁-C₄haloalkyl, C₂-C₄alkenyl, (C₅-C₇cycloalkyl)C₀-C₂alkyl, C₁-C₄alkoxy,C₂-C₄alkyl ether, C₂-C₄alkanoyl, C₁-C₄alkylsulfonyl, C₂-C₄alkanoylamino,mono- and di-(C₁-C₄alkyl)amino, mono- and di-(C₁-C₄alkyl)aminocarbonyl,mono- or di-(C₁-C₄alkyl)aminosulfonyl, phenylC₀-C₄alkyl and (4- to7-membered heterocycle)C₀-C₄alkyl, each of which is substituted withfrom 0 to 4 secondary substituents independently chosen from halogen,hydroxy, cyano, —COOH, C₁-C₄alkyl and C₁-C₄haloalkyl. Certain such R₃groups include azetidinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl,piperidinyl, piperazinyl, tetrahydropyridyl and azepanyl, each of whichis substituted with from 0 to 4 substituents independently chosen from:(a) halogen, hydroxy, amino, oxo, aminocarbonyl, aminosulfonyl and—COOH; and (b) C₁-C₄alkyl, C₂-C₄alkenyl, C₅-C₇cycloalkyl,C₁-C₄haloalkyl, C₁-C₄alkoxy, C₂-C₄alkyl ether, C₂-C₄alkanoyl,C₁-C₄alkylsulfonyl, C₂-C₄alkanoylamino and mono- anddi-(C₁-C₄alkyl)amino, each of which is substituted with from 0 to 4secondary substituents independently chosen from hydroxy and halogen.Representative examples of such R₃ groups include the heterocycles:

And substituted heterocycles, such as:

including enantiomers thereof

Other such R₃ groups include phenyl, pyridyl, pyrimidyl, pyrazinyl,pyridazinyl, imidazolyl, thienyl, oxazolyl and tetrahydrofuranyl, eachof which is substituted with from 0 to 4 substituents independentlychosen from halogen, hydroxy, amino, aminocarbonyl, aminosulfonyl,—COOH, C₁-C₄alkyl, C₅-C₇cycloalkyl, C₂-C₄alkyl ether, C₁-C₄alkoxy,C₂-C₄alkanoyl, C₁-C₄haloalkyl and mono- and di-(C₁-C₄alkyl)amino. Incertain embodiments, R₃ is not —NH₂. In other words, if R₃ has theformula:

and L is a single covalent bond, then at least one of R₅ and R₆ is nothydrogen.

In further embodiments, R₃ is

In certain such compounds, L is C₀-C₃alkylene; and R₇ is C₁-C₆alkyl,C₂-C₆alkenyl, (C₅-C₇cycloalkyl)C₀-C₄alkyl, C₂-C₄alkanoyl,phenylC₀-C₆alkyl or (6-membered heteroaryl)C₀-C₄alkyl, each of which issubstituted with from 0 to 4 substituents independently chosen fromhalogen, hydroxy, amino, aminocarbonyl, aminosulfonyl, —COOH,C₁-C₄alkyl, C₅-C₇cycloalkyl, C₂-C₄alkyl ether, C₁-C₄alkoxy,C₂-C₄alkanoyl, C₁-C₄haloalkyl and mono- and di-(C₁-C₄alkyl)amino. SuchR₃ groups include, for example, benzyloxy and C₁-C₆alkoxy, each of whichis optionally substituted with halogen, methyl, methoxy ortrifluoromethyl.

Within still further embodiments, R₃ is hydrogen, C₁-C₆alkyl or ahalogen.

Within further embodiments of the above Formulas, R₃ is C₁-C₄alkyl,C₃-C₇cycloalkyl or C₁-C₄haloalkyl, each of which is substituted withfrom 0 to 4 substituents independently chosen from halogen, hydroxy,cyano, oxo, aminocarbonyl, aminosulfonyl, —COOH, C₃-C₇cycloalkyl, phenyland 4- to 7-membered heterocycle.

-   R₄

R₄, in certain substituted biaryl piperazinyl-pyridine analoguesprovided herein, represents zero substituents or one methyl, ethyl oroxo group; in certain embodiments such a substituent is located adjacentto the nitrogen atom that is bound to the core, as shown below:

The carbon to which a methyl or ethyl group is attached is chiral incertain embodiments (e.g., as shown in Formula V, VI, VII or VIII), inwhich the group

respectively.

In other embodiments, R₄ represents a single oxo substituent.

-   X, Y and Z

X, Y and Z, as noted above, are independently CR_(x) or N, such that atleast one of X, Y and Z is N. In certain embodiments, each R_(x) isindependently selected from hydrogen, fluoro, methyl, amino and cyano;each R_(x) is independently selected from hydrogen, methyl, amino andcyano; each R_(x) is independently chosen from hydrogen and methyl; oreach R_(x) is hydrogen. In certain representative compounds, Z is N(e.g., X and Y are CH). In other compounds provided herein, X is N(e.g., Y and Z are CH). In further compounds Z and X are N, X and Y areN or Z and Y are N. In still further compounds, X, Y and Z are each N.

Representative substituted biaryl piperazinyl-pyridine analoguesprovided herein include, but are not limited to, those specificallydescribed in Examples 1-3. It will be apparent that the specificcompounds recited herein are representative only, and are not intendedto limit the scope of the present invention. Further, as noted above,all compounds provided herein may be present as a free base, apharmaceutically acceptable salt or other form, such as a hydrate.

Within certain aspects of the present invention, substituted biarylpiperazinyl-pyridine analogues provided herein detectably alter(modulate) VR1 activity, as determined using an in vitro VR1 functionalassay such as a calcium mobilization assay, dorsal root ganglion assayor in vivo pain relief assay. As an initial screen for such activity, aVR1 ligand binding assay may be used. References herein to a “VR1 ligandbinding assay” are intended to refer to a standard in vitro receptorbinding assay such as that provided in Example 5, and a “calciummobilization assay” (also referred to herein as a “signal transductionassay”) may be performed as described in Example 6. Briefly, to assessbinding to VR1, a competition assay may be performed in which a VR1preparation is incubated with labeled (e.g., ¹²⁵I or ³H) compound thatbinds to VR1 (e.g., a capsaicin receptor agonist such as RTX) andunlabeled test compound. Within the assays provided herein, the VR1 usedis preferably mammalian VR1, more preferably human or rat VR1. Thereceptor may be recombinantly expressed or naturally expressed. The VR1preparation may be, for example, a membrane preparation from HEK293 orCHO cells that recombinantly express human VR1. Incubation with acompound that detectably modulates vanilloid ligand binding to VR1results in a decrease or increase in the amount of label bound to theVR1 preparation, relative to the amount of label bound in the absence ofthe compound. This decrease or increase may be used to determine theK_(i) at VR1 as described herein. In general, compounds that decreasethe amount of label bound to the VR1 preparation within such an assayare preferred.

As noted above, certain substituted biaryl piperazinyl-pyridineanalogues are VR1 antagonists. IC₅₀ values for such compounds may bedetermined using a standard in vitro VR1-mediated calcium mobilizationassay, as provided in Example 6. Briefly, cells expressing capsaicinreceptor are contacted with a compound of interest and with an indicatorof intracellular calcium concentration (e.g., a membrane permeablecalcium sensitivity dye such as Fluo-3 or Fura-2 (both of which areavailable, for example, from Molecular Probes, Eugene, Oreg.), each ofwhich produce a fluorescent signal when bound to Ca⁺⁺). Such contact ispreferably carried out by one or more incubations of the cells in bufferor culture medium comprising either or both of the compound and theindicator in solution. Contact is maintained for an amount of timesufficient to allow the dye to enter the cells (e.g., 1-2 h). Cells arewashed or filtered to remove excess dye and are then contacted with avanilloid receptor agonist (e.g., capsaicin, RTX or olvanil), typicallyat a concentration equal to the EC₅₀ concentration, and a fluorescenceresponse is measured. When agonist-contacted cells are contacted with acompound that is a VR1 antagonist the fluorescence response is generallyreduced by at least 20%, preferably at least 50% and more preferably atleast 80%, as compared to cells that are contacted with the agonist inthe absence of test compound. The IC₅₀ for VR1 antagonists providedherein is preferably less than 1 micromolar, less than 100 nM, less than10 nM or less than 1 nM. In certain embodiments, VR1 antagonistsprovided herein exhibit no detectable agonist activity an in vitro assayof capsaicin receptor agonism at a concentration of compound equal tothe IC₅₀. Certain such antagonists exhibit no detectable agonistactivity an in vitro assay of capsaicin receptor agonism at aconcentration of compound that is 100-fold higher than the IC₅₀.

Other substituted biaryl piperazinyl-pyridine analogues may be capsaicinreceptor agonists. Capsaicin receptor agonist activity may generally bedetermined as described in Example 6. When cells are contacted with 1micromolar of a compound that is a VR1 agonist, the fluorescenceresponse is generally increased by an amount that is at least 30% of theincrease observed when cells are contacted with 100 nM capsaicin. TheEC₅₀ for VR1 agonists provided herein is preferably less than 1micromolar, less than 100 nM or less than 10 nM.

VR1 modulating activity may also, or alternatively, be assessed using acultured dorsal root ganglion assay as provided in Example 9 and/or anin vivo pain relief assay as provided in Example 10. VR1 modulatorsprovided herein preferably have a statistically significant specificeffect on VR1 activity within one or more of the functional assaysprovided in Examples 6 and 10, herein.

Within certain embodiments, VR1 modulators provided herein do notsubstantially modulate ligand binding to other cell surface receptors,such as EGF receptor tyrosine kinase or the nicotinic acetylcholinereceptor. In other words, such modulators do not substantially inhibitactivity of a cell surface receptor such as the human epidermal growthfactor (EGF) receptor tyrosine kinase or the nicotinic acetylcholinereceptor (e.g., the IC₅₀ or IC₄₀ at such a receptor is preferablygreater than 1 micromolar, and most preferably greater than 10micromolar). Preferably, a modulator does not detectably inhibit EGFreceptor activity or nicotinic acetylcholine receptor activity at aconcentration of 0.5 micromolar, 1 micromolar or more preferably 10micromolar. Assays for determining cell surface receptor activity arecommercially available, and include the tyrosine kinase assay kitsavailable from Panvera (Madison, Wis.).

In certain embodiments, preferred VR1 modulators are non-sedating. Inother words, a dose of VR1 modulator that is twice the minimum dosesufficient to provide analgesia in an animal model for determining painrelief (such as a model provided in Example 10, herein) causes onlytransient (i.e., lasting for no more than ½ the time that pain relieflasts) or preferably no statistically significant sedation in an animalmodel assay of sedation (using the method described by Fitzgerald et al.(1988) Toxicology 49(2-3):433-9). Preferably, a dose that is five timesthe minimum dose sufficient to provide analgesia does not producestatistically significant sedation. More preferably, a VR1 modulatorprovided herein does not produce sedation at intravenous doses of lessthan 25 mg/kg (preferably less than 10 mg/kg) or at oral doses of lessthan 140 mg/kg (preferably less than 50 mg/kg, more preferably less than30 mg/kg).

If desired, compounds provided herein may be evaluated for certainpharmacological properties including, but not limited to, oralbioavailability (preferred compounds are orally bioavailable to anextent allowing for therapeutically effective concentrations of thecompound to be achieved at oral doses of less than 140 mg/kg, preferablyless than 50 mg/kg, more preferably less than 30 mg/kg, even morepreferably less than 10 mg/kg, still more preferably less than 1 mg/kgand most preferably less than 0.1 mg/kg), toxicity (a preferred compoundis nontoxic when a therapeutically effective amount is administered to asubject), side effects (a preferred compound produces side effectscomparable to placebo when a therapeutically effective amount of thecompound is administered to a subject), serum protein binding and invitro and in vivo half-life (a preferred compound exhibits an in vivohalf-life allowing for Q.I.D. dosing, preferably T.I.D. dosing, morepreferably B.I.D. dosing, and most preferably once-a-day dosing). Inaddition, differential penetration of the blood brain barrier may bedesirable for VR1 modulators used to treat pain by modulating CNS VR1activity such that total daily oral doses as described above providesuch modulation to a therapeutically effective extent, while low brainlevels of VR1 modulators used to treat peripheral nerve mediated painmay be preferred (i.e., such doses do not provide brain (e.g., CSF)levels of the compound sufficient to significantly modulate VR1activity). Routine assays that are well known in the art may be used toassess these properties, and identify superior compounds for aparticular use. For example, assays used to predict bioavailabilityinclude transport across human intestinal cell monolayers, includingCaco-2 cell monolayers. Penetration of the blood brain barrier of acompound in humans may be predicted from the brain levels of thecompound in laboratory animals given the compound (e.g., intravenously).Serum protein binding may be predicted from albumin binding assays.Compound half-life is inversely proportional to the frequency of dosageof a compound. In vitro half-lives of compounds may be predicted fromassays of microsomal half-life as described within Example 7, herein.

As noted above, preferred compounds provided herein are nontoxic. Ingeneral, the term “nontoxic” shall be understood in a relative sense andis intended to refer to any substance that has been approved by theUnited States Food and Drug Administration (“FDA”) for administration tomammals (preferably humans) or, in keeping with established criteria, issusceptible to approval by the FDA for administration to mammals(preferably humans). In addition, a highly preferred nontoxic compoundgenerally satisfies one or more of the following criteria: (1) does notsubstantially inhibit cellular ATP production; (2) does notsignificantly prolong heart QT intervals; (3) does not cause substantialliver enlargement, or (4) does not cause substantial release of liverenzymes.

As used herein, a compound that does not substantially inhibit cellularATP production is a compound that satisfies the criteria set forth inExample 8, herein. In other words, cells treated as described in Example8 with 100 μM of such a compound exhibit ATP levels that are at least50% of the ATP levels detected in untreated cells. In more highlypreferred embodiments, such cells exhibit ATP levels that are at least80% of the ATP levels detected in untreated cells.

A compound that does not significantly prolong heart QT intervals is acompound that does not result in a statistically significantprolongation of heart QT intervals (as determined byelectrocardiography) in guinea pigs, minipigs or dogs uponadministration of a dose that yields a serum concentration equal to theEC₅₀ or IC₅₀ for the compound. In certain preferred embodiments, a doseof 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 40 or 50 mg/kg administeredparenterally or orally does not result in a statistically significantprolongation of heart QT intervals. By “statistically significant” ismeant results varying from control at the p<0.1 level or more preferablyat the p<0.05 level of significance as measured using a standardparametric assay of statistical significance such as a student's T test.

A compound does not cause substantial liver enlargement if dailytreatment of laboratory rodents (e.g., mice or rats) for 5-10 days witha dose that yields a serum concentration equal to the EC₅₀ or IC₅₀ forthe compound results in an increase in liver to body weight ratio thatis no more than 100% over matched controls. In more highly preferredembodiments, such doses do not cause liver enlargement of more than 75%or 50% over matched controls. If non-rodent mammals (e.g., dogs) areused, such doses should not result in an increase of liver to bodyweight ratio of more than 50%, preferably not more than 25%, and morepreferably not more than 10% over matched untreated controls. Preferreddoses within such assays include 0.01, 0.05. 0.1, 0.5, 1, 5, 10, 40 or50 mg/kg administered parenterally or orally.

Similarly, a compound does not promote substantial release of liverenzymes if administration of twice the minimum dose that yields a serumconcentration equal to the EC₅₀ or IC₅₀ at VR1 for the compound does notelevate serum levels of ALT, LDH or AST in laboratory rodents by morethan 100% over matched mock-treated controls. In more highly preferredembodiments, such doses do not elevate such serum levels by more than75% or 50% over matched controls. Alternatively, a compound does notpromote substantial release of liver enzymes if, in an in vitrohepatocyte assay, concentrations (in culture media or other suchsolutions that are contacted and incubated with hepatocytes in vitro)that are equal to the EC₅₀ or IC₅₀ for the compound do not causedetectable release of any of such liver enzymes into culture mediumabove baseline levels seen in media from matched mock-treated controlcells. In more highly preferred embodiments, there is no detectablerelease of any of such liver enzymes into culture medium above baselinelevels when such compound concentrations are five-fold, and preferablyten-fold the EC₅₀ or IC₅₀ for the compound.

In other embodiments, certain preferred compounds do not inhibit orinduce microsomal cytochrome P450 enzyme activities, such as CYP1A2activity, CYP2A6 activity, CYP2C9 activity, CYP2C19 activity, CYP2D6activity, CYP2E1 activity or CYP3A4 activity at a concentration equal tothe EC₅₀ or IC₅₀ at VR1 for the compound.

Certain preferred compounds are not clastogenic (e.g., as determinedusing a mouse erythrocyte precursor cell micronucleus assay, an Amesmicronucleus assay, a spiral micronucleus assay or the like) at aconcentration equal the EC₅₀ or IC₅₀ for the compound. In otherembodiments, certain preferred compounds do not induce sister chromatidexchange (e.g., in Chinese hamster ovary cells) at such concentrations.

For detection purposes, as discussed in more detail below, VR1modulators provided herein may be isotopically-labeled or radiolabeled.For example, compounds may have one or more atoms replaced by an atom ofthe same element having an atomic mass or mass number different from theatomic mass or mass number usually found in nature. Examples of isotopesthat can be present in the compounds provided herein include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and chlorine,such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S, ¹⁸F and³⁶Cl. In addition, substitution with heavy isotopes such as deuterium(i.e., ²H) can afford certain therapeutic advantages resulting fromgreater metabolic stability, for example increased in vivo half-life orreduced dosage requirements and, hence, may be preferred in somecircumstances.

Preparation of Substituted Biaryl Piperazinyl-Pyridine Analogues

Substituted biaryl piperazinyl-pyridine analogues may generally beprepared using standard synthetic methods. Starting materials arecommercially available from suppliers such as Sigma-Aldrich Corp. (St.Louis, Mo.), or may be synthesized from commercially availableprecursors using established protocols. By way of example, a syntheticroute similar to that shown in any of the following Schemes may be used,together with synthetic methods known in the art of synthetic organicchemistry. Each variable in the following schemes refers to any groupconsistent with the description of the compounds provided herein.

In the Schemes that follow the term “reduction” refers to the process ofreducing a nitro functionality to an amino functionality, the process oftransforming an ester functionality to an alcohol or the process oftransforming an amide to an amino group. The reduction of a nitro groupcan be carried out in a number of ways well known to those skilled inthe art of organic synthesis including, but not limited to, catalytichydrogenation, reduction with SnCl₂ and reduction with titaniumtrichloride. The reduction of an ester group is typically performedusing metal hydride reagents including, but not limited to,diisobutyl-aluminum hydride (DIBAL), lithium aluminum hydride (LAH), andsodium borohydride. The reduction of an amide can be carried outconveniently with reagents including, but not limited to, diborane aswell as lithium aluminum hydride (LAH). For an overview of reductionmethods see: Hudlicky, M. Reductions in Organic Chemistry, ACS Monograph188, 1996.

In the Schemes that follow, the term “hydrolyze” refers to the reactionof a substrate or reactant with water. More specifically, “hydrolyze”refers to the conversion of an ester or nitrile functionality into acarboxylic acid. This process can be catalyzed by a variety of acids orbases well known to those skilled in the art of organic synthesis.

In the Schemes that follow, the term “catalyst” refers to a suitabletransition metal catalyst such as, but not limited to,tetrakis(triphenylphosphine)palladium(0) or palladium(II) acetate. Inaddition, the catalytic systems may include ligands such as, but notlimited to, 2-(Dicyclohexylphosphino)biphenyl andtri-tert-butylphosphine, and may also include a base such as K₃PO₄,Na₂CO₃ or sodium or potassium tert-butoxide. Transition metal-catalyzedreactions can be carried out at ambient or elevated temperatures usingvarious inert solvents including, but not limited to, toluene, dioxane,DMF, N-methylpyrrolidinone, ethyleneglycol, dimethyl ether, diglyme andacetonitrile. When used in conjunction with suitable metallo-arylreagents, transition metal-catalyzed (hetero)aryl/aryl couplingreactions can be used to prepare the compounds encompassed in generalstructures 1D and 1E (Scheme 1), and 2C (Scheme 2), 4E (Scheme 4), 5B(Scheme 5), 6-F (Scheme 6), 8B and 8D (Scheme 8), 9C (Scheme 9), and 10C(Scheme 10). Commonly employed reagent/catalyst pairs include arylboronic acid/palladium(0) (Suzuki reaction; Miyaura and Suzuki (1995)Chemical Reviews 95:2457) and aryl trialkylstannane/palladium(0) (Stillereaction; T. N. Mitchell, (1992) Synthesis 9:803-815),arylzinc/palladium(0) and aryl Grignard/nickel(II). In addition,metal-catalyzed (hetero)aryl/amine coupling reactions (Buchwald-Hartwigcross-coupling reaction; J. F. Hartwig, Angew. Chem. Int. Ed.37:2046-2067 (1998)) can be used to prepare the compounds encompassed ingeneral structures 7F (Scheme 7), 9E (Scheme 9), and 10E (Scheme 10).The term “transmetalation,” as used in Schemes 2, 8, and 13, refers toone or more fo the above referenced metal-catalyzed cross couplingreactions which is suitable for coupling of (hetero)aryl halides withalcohols or amines. Certain transmetalation reaction conditions includethe Stille reaction and the Suzuki reaction.

In Schemes 11, 14 and 15, R₈ and R₉ are generally as described hereinfor R₅ and R₆ of Formula I. R₂₀, in Scheme 11, is any suitable leavinggroup, such as Br, Cl, I, OTf, mesylate or tosylate.

Certain definitions used in the following Schemes and elsewhere hereininclude:

-   BINAP (rac)-2,2′-Bis(diphenylphosphino)-1,1′-binaphthyl-   CDCl₃ deuterated chloroform-   δ chemical shift-   DCE 1,2-dichloroethane-   DCM dichloromethane or methylene chloride-   DDQ 2,3-dichloro-5,6-dicyano-1,4-benzoquinone-   DIBAL diisobutylaluminum hydride-   DIEA N,N-diisopropylethylamine-   DMA N,N-dimethylacetamide-   DMF dimethylformamide-   DMSO dimethylsulfoxide-   DPPF 1,1′-bis(diphenylphosphino)ferrocene-   Et₃N triethylamine-   EtOAc ethyl acetate-   EtOH ethanol-   h hour(s)-   ¹H NMR proton nuclear magnetic resonance-   HOAc acetic acid-   HPLC high pressure liquid chromatography-   Hz hertz-   KOAc potassium acetate-   LCMS liquid chromatography/mass spectrometry-   MS mass spectrometry-   (M+1) mass+1-   m-CPB A m-chloroperbenzoic acid-   MeOH methanol-   min minute(s)-   MsCl methanesulfonyl chloride-   NaNHCN sodium cyanamide-   n-BuLi n-butyl lithium-   t-Bu tertiary butyl-   Tf —SO₂CF₃-   Pd₂(dba)₃ tris(dibenzylideneacetone)dipalladium(0)-   Pd(PPh₃)₄ tetrakis(triphenylphosphine)palladium(0)-   PhNEt₂ diethyl-phenyl-amine, also referred to as N,N-diethylaniline-   PPh₃ triphenylphosphine-   Selectfluor®    1-Chloromethyl-4-Fluoro-1,4-Diazoniabicyclo[2.2.2]OctaneBis    (Tetrafluoroborate)-   t-BuOK potassium tert-butoxide-   TFA trifluoroacetic acid-   THF tetrahydrofuran-   TLC thin layer chromatography

In certain embodiments, a compound provided herein may contain one ormore asymmetric carbon atoms, so that the compound can exist indifferent stereoisomeric forms. Such forms can be, for example,racemates or optically active forms. As noted above, all stereoisomersare encompassed by the present invention. Nonetheless, it may bedesirable to obtain single enantiomers (i.e., optically active forms).Standard methods for preparing single enantiomers include asymmetricsynthesis and resolution of the racemates. Resolution of the racematescan be accomplished, for example, by conventional methods such ascrystallization in the presence of a resolving agent, or chromatographyusing, for example a chiral HPLC column.

Compounds may be radiolabeled by carrying out their synthesis usingprecursors comprising at least one atom that is a radioisotope. Eachradioisotope is preferably carbon (e.g., ¹⁴C), hydrogen (e.g., ³H),sulfur (e.g., ³⁵S) or iodine (e.g., ¹²⁵I). Tritium labeled compounds mayalso be prepared catalytically via platinum-catalyzed exchange intritiated acetic acid, acid-catalyzed exchange in tritiatedtrifluoroacetic acid, or heterogeneous-catalyzed exchange with tritiumgas using the compound as substrate. In addition, certain precursors maybe subjected to tritium-halogen exchange with tritium gas, tritium gasreduction of unsaturated bonds, or reduction using sodium borotritide,as appropriate. Preparation of radiolabeled compounds may beconveniently performed by a radioisotope supplier specializing in customsynthesis of radiolabeled probe compounds.

Pharmaceutical Compositions

The present invention also provides pharmaceutical compositionscomprising one or more compounds provided herein, together with at leastone physiologically acceptable carrier or excipient. Pharmaceuticalcompositions may comprise, for example, one or more of water, buffers(e.g., neutral buffered saline or phosphate buffered saline), ethanol,mineral oil, vegetable oil, dimethylsulfoxide, carbohydrates (e.g.,glucose, mannose, sucrose or dextrans), mannitol, proteins, adjuvants,polypeptides or amino acids such as glycine, antioxidants, chelatingagents such as EDTA or glutathione and/or preservatives. In addition,other active ingredients may (but need not) be included in thepharmaceutical compositions provided herein.

Pharmaceutical compositions may be formulated for any appropriate mannerof administration, including, for example, topical, oral, nasal, rectalor parenteral administration. The term parenteral as used hereinincludes subcutaneous, intradermal, intravascular (e.g., intravenous),intramuscular, spinal, intracranial, intrathecal and intraperitonealinjection, as well as any similar injection or infusion technique. Incertain embodiments, compositions suitable for oral use are preferred.Such compositions include, for example, tablets, troches, lozenges,aqueous or oily suspensions, dispersible powders or granules, emulsion,hard or soft capsules, or syrups or elixirs. Within yet otherembodiments, compositions of the present invention may be formulated asa lyophilizate. Formulation for topical administration may be preferredfor certain conditions (e.g., in the treatment of skin conditions suchas burns or itch). Formulation for direct administration into thebladder (intravesicular administration) may be preferred for treatmentof urinary incontinence and overactive bladder.

Compositions intended for oral use may further comprise one or morecomponents such as sweetening agents, flavoring agents, coloring agentsand/or preserving agents in order to provide appealing and palatablepreparations. Tablets contain the active ingredient in admixture withphysiologically acceptable excipients that are suitable for themanufacture of tablets. Such excipients include, for example, inertdiluents (e.g., calcium carbonate, sodium carbonate, lactose, calciumphosphate or sodium phosphate), granulating and disintegrating agents(e.g., corn starch or alginic acid), binding agents (e.g., starch,gelatin or acacia) and lubricating agents (e.g., magnesium stearate,stearic acid or talc). The tablets may be uncoated or they may be coatedby known techniques to delay disintegration and absorption in thegastrointestinal tract and thereby provide a sustained action over alonger period. For example, a time delay material such as glycerylmonosterate or glyceryl distearate may be employed.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent(e.g., calcium carbonate, calcium phosphate or kaolin), or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium (e.g., peanut oil, liquid paraffin or olive oil).

Aqueous suspensions contain the active material(s) in admixture withsuitable excipients, such as suspending agents (e.g., sodiumcarboxymethylcellulose, methylcellulose, hydropropylmethylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia);and dispersing or wetting agents (e.g., naturally-occurring phosphatidessuch as lecithin, condensation products of an alkylene oxide with fattyacids such as polyoxyethylene stearate, condensation products ofethylene oxide with long chain aliphatic alcohols such asheptadecaethyleneoxycetanol, condensation products of ethylene oxidewith partial esters derived from fatty acids and a hexitol such aspolyoxyethylene sorbitol monooleate, or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolanhydrides such as polyethylene sorbitan monooleate). Aqueoussuspensions may also comprise one or more preservatives, such as ethylor n-propyl p-hydroxybenzoate, one or more coloring agents, one or moreflavoring agents, and/or one or more sweetening agents, such as sucroseor saccharin.

Oily suspensions may be formulated by suspending the activeingredient(s) in a vegetable oil (e.g., arachis oil, olive oil, sesameoil or coconut oil) or in a mineral oil such as liquid paraffin. Theoily suspensions may contain a thickening agent such as beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and/or flavoring agents may be added to provide palatable oralpreparations. Such suspensions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, a suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, such as sweetening, flavoring and coloringagents, may also be present.

Pharmaceutical compositions may also be formulated as oil-in-wateremulsions. The oily phase may be a vegetable oil (e.g., olive oil orarachis oil), a mineral oil (e.g., liquid paraffin) or a mixturethereof. Suitable emulsifying agents include naturally-occurring gums(e.g., gum acacia or gum tragacanth), naturally-occurring phosphatides(e.g., soy bean lecithin, and esters or partial esters derived fromfatty acids and hexitol), anhydrides (e.g., sorbitan monoleate) andcondensation products of partial esters derived from fatty acids andhexitol with ethylene oxide (e.g., polyoxyethylene sorbitan monoleate).An emulsion may also comprise one or more sweetening and/or flavoringagents.

Syrups and elixirs may be formulated with sweetening agents, such asglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso comprise one or more demulcents, preservatives, flavoring agentsand/or coloring agents.

Formulations for topical administration typically comprise a topicalvehicle combined with active agent(s), with or without additionaloptional components. Suitable topical vehicles and additional componentsare well known in the art, and it will be apparent that the choice of avehicle will depend on the particular physical form and mode ofdelivery. Topical vehicles include water; organic solvents such asalcohols (e.g., ethanol or isopropyl alcohol) or glycerin; glycols(e.g., butylene, isoprene or propylene glycol); aliphatic alcohols(e.g., lanolin); mixtures of water and organic solvents and mixtures oforganic solvents such as alcohol and glycerin; lipid-based materialssuch as fatty acids, acylglycerols (including oils, such as mineral oil,and fats of natural or synthetic origin), phosphoglycerides,sphingolipids and waxes; protein-based materials such as collagen andgelatin; silicone-based materials (both non-volatile and volatile); andhydrocarbon-based materials such as microsponges and polymer matrices. Acomposition may further include one or more components adapted toimprove the stability or effectiveness of the applied formulation, suchas stabilizing agents, suspending agents, emulsifying agents, viscosityadjusters, gelling agents, preservatives, antioxidants, skin penetrationenhancers, moisturizers and sustained release materials. Examples ofsuch components are described in Martindale—The Extra Pharmacopoeia(Pharmaceutical Press, London 1993) and Martin (ed.), Remington'sPharmaceutical Sciences. Formulations may comprise microcapsules, suchas hydroxymethylcellulose or gelatin-microcapsules, liposomes, albuminmicrospheres, microemulsions, nanoparticles or nanocapsules.

A topical formulation may be prepared in any of a variety of physicalforms including, for example, solids, pastes, creams, foams, lotions,gels, powders, aqueous liquids and emulsions. The physical appearanceand viscosity of such pharmaceutically acceptable forms can be governedby the presence and amount of emulsifier(s) and viscosity adjuster(s)present in the formulation. Solids are generally firm and non-pourableand commonly are formulated as bars or sticks, or in particulate form;solids can be opaque or transparent, and optionally can containsolvents, emulsifiers, moisturizers, emollients, fragrances,dyes/colorants, preservatives and other active ingredients that increaseor enhance the efficacy of the final product. Creams and lotions areoften similar to one another, differing mainly in their viscosity; bothlotions and creams may be opaque, translucent or clear and often containemulsifiers, solvents, and viscosity adjusting agents, as well asmoisturizers, emollients, fragrances, dyes/colorants, preservatives andother active ingredients that increase or enhance the efficacy of thefinal product. Gels can be prepared with a range of viscosities, fromthick or high viscosity to thin or low viscosity. These formulations,like those of lotions and creams, may also contain solvents,emulsifiers, moisturizers, emollients, fragrances, dyes/colorants,preservatives and other active ingredients that increase or enhance theefficacy of the final product. Liquids are thinner than creams, lotions,or gels and often do not contain emulsifiers. Liquid topical productsoften contain solvents, emulsifiers, moisturizers, emollients,fragrances, dyes/colorants, preservatives and other active ingredientsthat increase or enhance the efficacy of the final product.

Suitable emulsifiers for use in topical formulations include, but arenot limited to, ionic emulsifiers, cetearyl alcohol, non-ionicemulsifiers like polyoxyethylene oleyl ether, PEG-40 stearate,ceteareth-12, ceteareth-20, ceteareth-30, ceteareth alcohol, PEG-100stearate and glyceryl stearate. Suitable viscosity adjusting agentsinclude, but are not limited to, protective colloids or non-ionic gumssuch as hydroxyethylcellulose, xanthan gum, magnesium aluminum silicate,silica, microcrystalline wax, beeswax, paraffin, and cetyl palmitate. Agel composition may be formed by the addition of a gelling agent such aschitosan, methyl cellulose, ethyl cellulose, polyvinyl alcohol,polyquaterniums, hydroxyethylcellulose, hydroxypropylcellulose,hydroxypropylmethylcellulose, carbomer or ammoniated glycyrrhizinate.Suitable surfactants include, but are not limited to, nonionic,amphoteric, ionic and anionic surfactants. For example, one or more ofdimethicone copolyol, polysorbate 20, polysorbate 40, polysorbate 60,polysorbate 80, lauramide DEA, cocamide DEA, and cocamide MEA, oleylbetaine, cocamidopropyl phosphatidyl PG-dimonium chloride, and ammoniumlaureth sulfate may be used within topical formulations. Suitablepreservatives include, but are not limited to, antimicrobials such asmethylparaben, propylparaben, sorbic acid, benzoic acid, andformaldehyde, as well as physical stabilizers and antioxidants such asvitamin E, sodium ascorbate/ascorbic acid and propyl gallate. Suitablemoisturizers include, but are not limited to, lactic acid and otherhydroxy acids and their salts, glycerin, propylene glycol, and butyleneglycol. Suitable emollients include lanolin alcohol, lanolin, lanolinderivatives, cholesterol, petrolatum, isostearyl neopentanoate andmineral oils. Suitable fragrances and colors include, but are notlimited to, FD&C Red No. 40 and FD&C Yellow No. 5. Other suitableadditional ingredients that may be included a topical formulationinclude, but are not limited to, abrasives, absorbents, anti-cakingagents, anti-foaming agents, anti-static agents, astringents (e.g.,witch hazel, alcohol and herbal extracts such as chamomile extract),binders/excipients, buffering agents, chelating agents, film formingagents, conditioning agents, propellants, opacifying agents, pHadjusters and protectants.

An example of a suitable topical vehicle for formulation of a gel is:hydroxypropylcellulose (2.1%); 70/30 isopropyl alcohol/water (90.9%);propylene glycol (5.1%); and Polysorbate 80 (1.9%). An example of asuitable topical vehicle for formulation as a foam is: cetyl alcohol(1.1%); stearyl alcohol (0.5%; Quaternium 52 (1.0%); propylene glycol(2.0%); Ethanol 95 PGF3 (61.05%); deionized water (30.05%); P75hydrocarbon propellant (4.30%). All percents are by weight.

Typical modes of delivery for topical compositions include applicationusing the fingers; application using a physical applicator such as acloth, tissue, swab, stick or brush; spraying (including mist, aerosolor foam spraying); dropper application; sprinkling; soaking; andrinsing. Controlled release vehicles can also be used.

A pharmaceutical composition may be prepared as a sterile injectibleaqueous or oleaginous suspension. The compound(s) provided herein,depending on the vehicle and concentration used, can either be suspendedor dissolved in the vehicle. Such a composition may be formulatedaccording to the known art using suitable dispersing, wetting agentsand/or suspending agents such as those mentioned above. Among theacceptable vehicles and solvents that may be employed are water,1,3-butanediol, Ringer's solution and isotonic sodium chloride solution.In addition, sterile, fixed oils may be employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employed,including synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectible compositions,and adjuvants such as local anesthetics, preservatives and/or bufferingagents can be dissolved in the vehicle.

Pharmaceutical compositions may also be formulated as suppositories(e.g., for rectal administration). Such compositions can be prepared bymixing the drug with a suitable non-irritating excipient that is solidat ordinary temperatures but liquid at the rectal temperature and willtherefore melt in the rectum to release the drug. Suitable excipientsinclude, for example, cocoa butter and polyethylene glycols.

Pharmaceutical compositions may be formulated as sustained release orcontrolled-release formulations (i.e., a formulation such as a capsulethat effects a slow release of modulator following administration). Suchformulations may generally be prepared using well known technology andadministered by, for example, oral, rectal or subcutaneous implantation,or by implantation at the desired target site. Carriers for use withinsuch formulations are biocompatible, and may also be biodegradable;preferably the formulation provides a relatively constant level ofmodulator release. The amount of modulator contained within a sustainedrelease formulation depends upon, for example, the site of implantation,the rate and expected duration of release and the nature of thecondition to be treated or prevented.

In addition to or together with the above modes of administration, acompound provided herein may be conveniently added to food or drinkingwater (e.g., for administration to non-human animals including companionanimals (such as dogs and cats) and livestock). Animal feed and drinkingwater compositions may be formulated so that the animal takes in anappropriate quantity of the composition along with its diet. It may alsobe convenient to present the composition as a premix for addition tofeed or drinking water.

Compounds are generally administered in a therapeutically effectiveamount. Preferred systemic doses are no higher than 50 mg per kilogramof body weight per day (e.g., ranging from about 0.001 mg to about 50 mgper kilogram of body weight per day), with oral doses generally beingabout 5-20 fold higher than intravenous doses (e.g., ranging from 0.01to 40 mg per kilogram of body weight per day).

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage unit will vary depending, forexample, upon the patient being treated and the particular mode ofadministration. Dosage units will generally contain from about 10 μg toabout 500 mg of an active ingredient. Optimal dosages may be establishedusing routine testing, and procedures that are well known in the art.

Pharmaceutical compositions may be packaged for treating conditionsresponsive to VR1 modulation (e.g., treatment of exposure to vanilloidligand or other irritant, pain, itch, obesity or urinary incontinence).Packaged pharmaceutical compositions generally include (i) a containerholding a pharmaceutical composition that comprises at least one VR1modulator as described herein and (ii) instructions (e.g., labeling or apackage insert) indicating that the contained composition is to be usedfor treating a condition responsive to VR1 modulation in the patient.

Methods of Use

VR1 modulators provided herein may be used to alter activity and/oractivation of capsaicin receptors in a variety of contexts, both invitro and in vivo. Within certain aspects, VR1 antagonists may be usedto inhibit the binding of vanilloid ligand agonist (such as capsaicinand/or RTX) to capsaicin receptor in vitro or in vivo. In general, suchmethods comprise the step of contacting a capsaicin receptor with one ormore VR1 modulators provided herein, in the presence of vanilloid ligandin aqueous solution and under conditions otherwise suitable for bindingof the ligand to capsaicin receptor. The VR1 modulator(s) are generallypresent at a concentration that is sufficient to alter the binding ofvanilloid ligand to VR1 in vitro (using the assay provided in Example 5)and/or VR1-mediated signal transduction (using an assay provided inExample 6). The capsaicin receptor may be present in solution orsuspension (e.g., in an isolated membrane or cell preparation), or in acultured or isolated cell. Within certain embodiments, the capsaicinreceptor is expressed by a neuronal cell present in a patient, and theaqueous solution is a body fluid. Preferably, one or more VR1 modulatorsare administered to an animal in an amount such that the VR1 modulatoris present in at least one body fluid of the animal at a therapeuticallyeffective concentration that is 1 micromolar or less; preferably 500nanomolar or less; more preferably 100 nanomolar or less, 50 nanomolaror less, 20 nanomolar or less, or 10 nanomolar or less. For example,such compounds may be administered at a therapeutically effective dosethat is less than 20 mg/kg body weight, preferably less than 5 mg/kgand, in some instances, less than 1 mg/kg.

Also provided herein are methods for modulating, preferably reducing,the signal-transducing activity (i.e., the calcium conductance) of acellular capsaicin receptor. Such modulation may be achieved bycontacting a capsaicin receptor (either in vitro or in vivo) with one ormore VR1 modulators provided herein under conditions suitable forbinding of the modulator(s) to the receptor. The VR1 modulator(s) aregenerally present at a concentration that is sufficient to alter thebinding of vanilloid ligand to VR1 in vitro and/or VR1-mediated signaltransduction as described herein. The receptor may be present insolution or suspension, in a cultured or isolated cell preparation or ina cell within a patient. For example, the cell may be a neuronal cellthat is contacted in vivo in an animal. Alternatively, the cell may bean epithelial cell, such as a urinary bladder epithelial cell(urothelial cell) or an airway epithelial cell that is contacted in vivoin an animal. Modulation of signal tranducing activity may be assessedby detecting an effect on calcium ion conductance (also referred to ascalcium mobilization or flux). Modulation of signal transducing activitymay alternatively be assessed by detecting an alteration of a symptom(e.g., pain, burning sensation, broncho-constriction, inflammation,cough, hiccup, itch, urinary incontinence or overactive bladder) of apatient being treated with one or more VR1 modulators provided herein.

VR1 modulator(s) provided herein are preferably administered to apatient (e.g., a human) orally or topically, and are present within atleast one body fluid of the animal while modulating VR1signal-transducing activity. Preferred VR1 modulators for use in suchmethods modulate VR1 signal-transducing activity in vitro at aconcentration of 1 nanomolar or less, preferably 100 picomolar or less,more preferably 20 picomolar or less, and in vivo at a concentration of1 micromolar or less, 500 nanomolar or less, or 100 nanomolar or less ina body fluid such as blood.

The present invention further provides methods for treating conditionsresponsive to VR1 modulation. Within the context of the presentinvention, the term “treatment” encompasses both disease-modifyingtreatment and symptomatic treatment, either of which may be prophylactic(i.e., before the onset of symptoms, in order to prevent, delay orreduce the severity of symptoms) or therapeutic (i.e., after the onsetof symptoms, in order to reduce the severity and/or duration ofsymptoms). A condition is “responsive to VR1 modulation” if it ischaracterized by inappropriate activity of a capsaicin receptor,regardless of the amount of vanilloid ligand present locally, and/or ifmodulation of capsaicin receptor activity results in alleviation of thecondition or a symptom thereof. Such conditions include, for example,symptoms resulting from exposure to VR1-activating stimuli, pain,respiratory disorders (such as cough, asthma, chronic obstructivepulmonary disease, chronic bronchitis, cystic fibrosis and rhinitis,including allergic rhinitis, such as seasonal an perennial rhinitis, andnon-allergic rhinitis) itch, urinary incontinence, overactive bladder,hiccup and obesity, as described in more detail below. Such conditionsmay be diagnosed and monitored using criteria that have been establishedin the art. Patients may include humans, domesticated companion animalsand livestock, with dosages as described above.

Treatment regimens may vary depending on the compound used and theparticular condition to be treated; however, for treatment of mostdisorders, a frequency of administration of 4 times daily or less ispreferred. In general, a dosage regimen of 2 times daily is morepreferred, with once a day dosing particularly preferred. For thetreatment of acute pain, a single dose that rapidly reaches effectiveconcentrations is desirable. It will be understood, however, that thespecific dose level and treatment regimen for any particular patientwill depend upon a variety of factors including the activity of thespecific compound employed, the age, body weight, general health, sex,diet, time of administration, route of administration, and rate ofexcretion, drug combination and the severity of the particular diseaseundergoing therapy. In general, the use of the minimum dose sufficientto provide effective therapy is preferred. Patients may generally bemonitored for therapeutic effectiveness using medical or veterinarycriteria suitable for the condition being treated or prevented.

Patients experiencing symptoms resulting from exposure to capsaicinreceptor-activating stimuli include individuals with burns caused byheat, light, tear gas or acid and those whose mucous membranes areexposed (e.g., via ingestion, inhalation or eye contact) to capsaicin(e.g., from hot peppers or in pepper spray) or a related irritant suchas acid, tear gas, infectious agent(s) or air pollutant(s). Theresulting symptoms (which may be treated using VR1 modulators,especially antagonists, provided herein) may include, for example, pain,broncho-constriction and inflammation.

Pain that may be treated using the VR1 modulators provided herein may bechronic or acute and includes, but is not limited to, peripheralnerve-mediated pain (especially neuropathic pain). Compounds providedherein may be used in the treatment of, for example, postmastectomy painsyndrome, stump pain, phantom limb pain, oral neuropathic pain,toothache (dental pain), denture pain, postherpetic neuralgia, diabeticneuropathy, chemotherapy-induced neuropathy, reflex sympatheticdystrophy, trigeminal neuralgia, osteoarthritis, rheumatoid arthritis,fibromyalgia, Guillain-Barre syndrome, meralgia paresthetica,burning-mouth syndrome and/or pain associated with nerve and rootdamage, including as pain associated with peripheral nerve disorders(e.g., nerve entrapment and brachial plexus avulsions, amputation,peripheral neuropathies including bilateral peripheral neuropathy, ticdouloureux, atypical facial pain, nerve root damage, and arachnoiditis).Additional neuropathic pain conditions include causalgia (reflexsympathetic dystrophy—RSD, secondary to injury of a peripheral nerve),neuritis (including, for example, sciatic neuritis, peripheral neuritis,polyneuritis, optic neuritis, postfebrile neuritis, migrating neuritis,segmental neuritis and Gombault's neuritis), neuronitis, neuralgias(e.g., those mentioned above, cervicobrachial neuralgia, cranialneuralgia, geniculate neuralgia, glossopharyngial neuralgia, migranousneuralgia, idiopathic neuralgia, intercostals neuralgia, mammaryneuralgia, mandibular joint neuralgia, Morton's neuralgia, nasociliaryneuralgia, occipital neuralgia, red neuralgia, Sluder's neuralgia,splenopalatine neuralgia, supraorbital neuralgia and vidian neuralgia),surgery-related pain, musculoskeletal pain, myofascial pain syndromes,AIDS-related neuropathy, MS-related neuropathy, central nervous systempain (e.g., pain due to brain stem damage, sciatica, and ankylosingspondylitis), and spinal pain, including spinal cord injury-relatedpain. Headache, including headaches involving peripheral nerve activitymay also be treated as described herein. Such pain includes, forexample, such as sinus, cluster (i.e., migranous neuralgia) and tensionheadaches, migraine, temporomandibular pain and maxillary sinus pain.For example, migraine headaches may be prevented by administration of acompound provided herein as soon as a pre-migrainous aura is experiencedby the patient. Further conditions that can be treated as describedherein include Charcot's pains, intestinal gas pains, ear pain, heartpain, muscle pain, eye pain, orofacial pain (e.g., odontalgia),abdominal pain, gynaecological pain (e.g., menstrual pain,dysmenorrhoea, pain associated with cystitis, labor pain, chronic pelvicpain, chronic prostitis and endometriosis), acute and chronic back pain(e.g., lower back pain), gout, scar pain, hemorrhoidal pain, dyspepticpains, angina, nerve root pain, “non-painful” neuropathies, complexregional pain syndrome, homotopic pain and heterotopic pain—includingpain associated with carcimnoma, often referred to as cancer pain (e.g.,in patients with bone cancer), pain (and inflammation) associated withvenom exposure (e.g., due to snake bite, spider bite, or insect sting)and trauma associated pain (e.g., post-surgical pain, episiotomy pain,pain from cuts, musculoskeletal pain, bruises and broken bones, and burnpain, especially primary hyperalgesia associated therewith). Additionalpain conditions that may be treated as described herein include painassociated with respiratory disorders as described above, autoimmunediseases, immunodeficiency disorders, hot flashes, inflammatory boweldisease, irritable bowel syndrome and/or inflammatory bowel disease. VR1modulators may also be used to treat depression and gastroesophagealreflux disease (GERD), including the pain associated with GERD.

Within certain aspects, VR1 modulators provided herein may be used forthe treatment of mechanical pain. As used herein, the term “mechanicalpain” refers to pain other than headache pain that is not neuropathic ora result of exposure to heat, cold or external chemical stimuli.Mechanical pain includes physical trauma (other than thermal or chemicalburns or other irritating and/or painful exposures to noxious chemicals)such as post-surgical pain and pain from cuts, bruises and broken bones;toothache; denture pain; nerve root pain; osteoartiritis; rheumatoidarthritis; fibromyalgia; meralgia paresthetica; back pain;cancer-associated pain; angina; carpel tunnel syndrome; and painresulting from bone fracture, labor, hemorrhoids, intestinal gas,dyspepsia, and menstruation.

Itching conditions that may be treated include psoriatic pruritus, itchdue to hemodialysis, aguagenic pruritus, and itching associated withvulvar vestibulitis, contact dermatitis, insect bites and skinallergies. Urinary tract conditions that may be treated as describedherein include urinary incontinence (including overflow incontinence,urge incontinence and stress incontinence), as well as overactive orunstable bladder conditions (including bladder detrusor hyper-reflexia,detrusor hyper-reflexia of spinal origin and bladder hypersensitivity).In certain such treatment methods, VR1 modulator is administered via acatheter or similar device, resulting in direct injection of VR1modulator into the bladder. Compounds provided herein may also be usedas anti-tussive agents (to prevent, relieve or suppress coughing) andfor the treatment of hiccup, and to promote weight loss in an obesepatient.

Within other aspects, VR1 modulators provided herein may be used withincombination therapy for the treatment of conditions involving painand/or inflammatory components. Such conditions include, for example,autoimmune disorders and pathologic autoimmune responses known to havean inflammatory component including, but not limited to, arthritis(especially rheumatoid arthritis), psoriasis, Crohn's disease, lupuserythematosus, irritable bowel syndrome, tissue graft rejection, andhyperacute rejection of transplanted organs. Other such conditionsinclude trauma (e.g., injury to the head or spinal cord), cardio- andcerebo-vascular disease and certain infectious diseases.

Within such combination therapy, a VR1 modulator is administered to apatient along with an analgesic and/or anti-inflammatory agent. The VR1modulator and analgesic and/or anti-inflammatory agent may be present inthe same pharmaceutical composition, or may be administered separatelyin either order. Anti-inflammatory agents include, for example,non-steroidal anti-inflammatory drugs (NSAIDs), non-specific andcyclooxygenase-2 (COX-2) specific cyclooxygenase enzyme inhibitors, goldcompounds, corticosteroids, methotrexate, tumor necrosis factor (TNF)receptor antagonists, anti-TNF alpha antibodies, anti-C5 antibodies, andinterleukin-1 (IL-1) receptor antagonists. Examples of NSAIDs include,but are not limited to ibuprofen (e.g., ADVIL™, MOTRIN™), flurbiprofen(ANSAID™), naproxen or naproxen sodium (e.g., NAPROSYN, ANAPROX,ALEVE™), diclofenac (e.g., CATAFLAM™, VOLTAREN™), combinations ofdiclofenac sodium and misoprostol (e.g., ARTHROTEC™), sulindac(CLINORIL™), oxaprozin (DAYPRO™), diflunisal (DOLOBID™), piroxicam(FELDENE™) indomethacin (INDOCIN™), etodolac (LODINE™), fenoprofencalcium (NALFON™) ketoprofen (e.g., ORUDIS™, ORUVAIL™), sodiumnabumetone (RELAFEN™), sulfasalazine (AZULFIDINE™), tolmetin sodium(TOLECTIN™), and hydroxychloroquine (PLAQUENIL™) One class of NSAIDsconsists of compounds that inhibit cyclooxygenase (COX) enzymes. NSAIDsfurther include salicylates such as acetylsalicylic acid or aspirin,sodium salicylate, choline and magnesium salicylates (TRILISATE™), andsalsalate (DISALCID™), as well as corticosteroids such as cortisone(CORTONE™ acetate), dexamethasone (e.g., DECADRON™) methylprednisolone(MEDROL™) prednisolone (PRELONE™), prednisolone sodium phosphate(PEDIAPRED™), and prednisone (e.g., PREDNICEN-M™, DELTASONE™,STERAPRED™). Further anti-inflammatoryr agents include meloxicam,rofecoxib, celecoxib, etoricoxib, parecoxib, valdecoxib and tilicoxib.

Suitable dosages for VR1 modulator within such combination therapy aregenerally as described above. Dosages and methods of administration ofanti-inflammatory agents can be found, for example, in themanufacturer's instructions in the Physician's Desk Reference. Incertain embodiments, the combination administration of a VR1 modulatorwith an anti-inflammatory agent results in a reduction of the dosage ofthe anti-inflammatory agent required to produce a therapeutic effect(i.e., a decrease in the minimum therapeutically effective amount).Thus, preferably, the dosage of anti-inflammatory agent in a combinationor combination treatment method of the invention is less than themaximum dose advised by the manufacturer for administration of theanti-inflammatory agent without combination administration of a VR1antagonist. More preferably this dosage is less than ¾, even morepreferably less than ½, and highly preferably, less than ¼ of themaximum dose, while most preferably the dose is less than 10% of themaximum dose advised by the manufacturer for administration of theanti-inflammatory agent(s) when administered without combinationadministration of a VR1 antagonist. It will be apparent that the dosageamount of VR1 antagonist component of the combination needed to achievethe desired effect may similarly be affected by the dosage amount andpotency of the anti-inflammatory agent component of the combination.

In certain preferred embodiments, the combination administration of aVR1 modulator with an anti-inflammatory agent is accomplished bypackaging one or more VR1 modulators and one or more anti-inflammatoryagents in the same package, either in separate containers within thepackage or in the same contained as a mixture of one or more VR1antagonists and one or more anti-inflammatory agents. Preferred mixturesare formulated for oral administration (e.g., as pills, capsules,tablets or the like). In certain embodiments, the package comprises alabel bearing indicia indicating that the one or more VR1 modulators andone or more anti-inflammatory agents are to be taken together for thetreatment of an inflammatory pain condition.

Within further aspects, VR1 modulators provided herein may be used incombination with one or more additional pain relief medications. Certainsuch medications are also anti-inflammatory agents, and are listedabove. Other such medications are analgesic agents, including narcoticagents which typically act at one or more opioid receptor subtypes(e.g., μ, κ and/or δ), preferably as agonists or partial agonists. Suchagents include opiates, opiate derivatives and opioids, as well aspharmaceutically acceptable salts and hydrates thereof. Specificexamples of narcotic analgesics include, within preferred embodiments,alfentanil, alphaprodine, anileridine, bezitramide, buprenorphine,butorphanol, codeine, diacetyldihydromorphine, diacetylmorphine,dihydrocodeine, diphenoxylate, ethylmorphine, fentanyl, heroin,hydrocodone, hydromorphone, isomethadone, levomethorphan, levorphane,levorphanol, meperidine, metazocine, methadone, methorphan, metopon,morphine, nalbuphine, opium extracts, opium fluid extracts, powderedopium, granulated opium, raw opium, tincture of opium, oxycodone,oxymorphone, paregoric, pentazocine, pethidine, phenazocine, piminodine,propoxyphene, racemethorphan, racemorphan, sulfentanyl, thebaine andpharmaceutically acceptable salts and hydrates of the foregoing agents.

Other examples of narcotic analgesic agents include acetorphine,acetyldihydrocodeine, acetylmethadol, allylprodine, alphracetylmethadol,alphameprodine, alphamethadol, benzethidine, benzylmorphine,betacetylmethadol, betameprodine, betamethadol, betaprodine,clonitazene, codeine methylbromide, codeine-N-oxide, cyprenorphine,desomorphine, dextromoramide, diampromide, diethylthiambutene,dihydromorphine, dimenoxadol, dimepheptanol, dimethylthiamubutene,dioxaphetyl butyrate, dipipanone, drotebanol, ethanol,ethylmethylthiambutene, etonitazene, etorphine, etoxeridine,furethidine, hydromorphinol, hydroxypethidine, ketobemidone,levomoramide, levophenacylmorphan, methyldesorphine,methyldihydromorphine, morpheridine, morphine methylpromide, morphinemethylsulfonate, morphine-N-oxide, myrophin, naloxone, naltyhexone,nicocodeine, nicomorphine, noracymethadol, norlevorphanol, normethadone,normorphine, norpipanone, pentazocaine, phenadoxone, phenampromide,phenomorphan, phenoperidine, piritramide, pholcodine, proheptazoine,properidine, propiran, racemoramide, thebacon, trimeperidine and thepharmaceutically acceptable salts and hydrates thereof.

Further specific representative analgesic agents include, for exampleacetaminophen (paracetamol); aspirin and other NSAIDs described above;NR2B antagonists; bradykinin antagonists; anti-migraine agents;anticonvulsants such as oxcarbazepine and carbamazepine; antidepressants(such as TCAs, SSRIs, SNR1s, substance P antagonists, etc.); spinalblocks; gabapentin; asthma treatments (such as β₂-adrenergic receptoragonists; leukotriene D₄ antagonists (e.g., montelukast); TALWIN® Nx andDEMEROL® (both available from Sanofi Winthrop Pharmaceuticals; New York,N.Y.); LEVO-DROMORAN®; BUPRENEX® (Reckitt & Coleman Pharmaceuticals,Inc.; Richmond, Va.); MSIR® (Purdue Pharma L.P.; Norwalk, Conn.);DILAUDID® (Knoll Pharmaceutical Co.; Mount Olive, N.J.); SUBLIMAZE®;SUFENTA®(Janssen Pharmaceutica Inc.; Titusville, N.J.); PERCOCET®,NUBAIN® and NUMORPHAN® (all available from Endo Pharmaceuticals Inc.;Chadds Ford, Pa.) HYDROSTAT® IR, MS/S and MS/L (all available fromRichwood Pharmaceutical Co. Inc; Florence, Ky.), ORAMORPH® SR andROXICODONE® (both available from Roxanne Laboratories; Columbus Ohio)and STADOL® (Bristol-Myers Squibb; New York, N.Y.). Still furtheranalgesic agents include CB2-receptor agonists, such as AM1241, andcompounds that bind to the α2δ subunit, such as Neurontin (Gabapentin)and pregabalin.

Representative anti-migraine agents for use in combination with a VR1modulator provided herein include CGRP antagonists, ergotamines and5-HT₁ agonists, such as sumatripan, naratriptan, zolmatriptan andrizatriptan.

Within still further aspects, VR1 modulators provided herein may be usedin combination with one or more leukotriene receptor antagonists (e.g.,agents that inhibits the cysteinyl leukotriene CysLT₁ receptor). CysLT₁antagonists include Montelukast (SINGULAIR®; Merck & Co., Inc.). Suchcombinations find use in the treatment of pulmonary disorders such asasthma.

For the treatment or prevention of cough, a VR1 modulator as providedherein may be used in combination with other medication designed totreat this condition, such as antibiotics, anti-inflammatory agents,cystinyl leukotrienes, histamine antagonists, corticosteroids, opioids,NMDA antagonists, proton pump inhibitors, nociceptin, neurokinin (NK1,NK2 and NK3) and bradykinin (BK1 and BK2) receptor antagonists,cannabinoids, blockers of Na+-dependent channels and large conductanceCa⁺²-dependent K⁺-channel activators. Specific agents includedexbrompheniramine plus pseudoephedrine, loratadine, oxymetazoline,ipratropium, albuterol, beclomethasone, morphine, codeine, pholcodeineand dextromethorphan.

The present invention further provides combination therapy for thetreatment of urinary incontinence. Within such aspects, a VR1 modulatorprovided herein may be used in combination with other medicationdesigned to treat this condition, such as estrogen replacement therapy,progesterone congeners, electrical stimulation, calcium channelblockers, antispasmodic agents, cholinergic antagonists, antimuscarinicdrugs, tricyclic antidepressants, SNRIs, beta adrenoceptor agonists,phosphodiesterase inhibitors, potassium channel openers,nociceptin/orphanin FQ (OP4) agonists, neurokinin (NK1 and NK2)antagonists, P2X3 antagonists, musculotrophic drugs and sacralneuromodulation. Specific agents include oxybutinin, emepronium,tolterodine, flavoxate, flurbiprofen, dicyclomine, propiverine,propantheline, imipramine, doxepin, duloxetine, 1-deamino-8-D-argininevasopressin, muscarinic receptor antagonists such as Tolterodine(DETROL®; Pharmacia Corporation) and anticholinergic agents such asOxybutynin (DITROPAN®; Ortho-McNeil Pharmaceutical, Inc., Raritan,N.J.).

Suitable dosages for VR1 modulator within such combination therapy aregenerally as described above. Dosages and methods of administration ofother pain relief medications can be found, for example, in themanufacturer's instructions in the Physician's Desk Reference. Incertain embodiments, the combination administration of a VR1 modulatorwith one or more additional pain medications results in a reduction ofthe dosage of each therapeutic agent required to produce a therapeuticeffect (e.g., the dosage or one or both agent may less than ¾, less than½, less than ¼ or less than 10% of the maximum dose listed above oradvised by the manufacturer).

For use in combination therapy, pharmaceutical compositions as describedabove may further comprise one or more additional medications asdescribed above. In certain such compositions, the additional medicationis an analgesic. Also provided herein are packaged pharmaceuticalpreparations comprising one or more VR1 modulators and one or moreadditional medications (e.g., analgesics) in the same package. Suchpackaged pharmaceutical preparations generally include (i) a containerholding a pharmaceutical composition that comprises at least one VR1modulator as described herein; (ii) a container holding a pharmaceuticalcomposition that comprises at least one additional medication (such as apain relief and/or anti-inflammatory medication) as described above and(iii) instructions (e.g., labeling or a package insert) indicating thatthe compositions are to be used simultaneously, separately orsequentially for treating or preventing a condition responsive to VR1modulation in the patient (such as a condition in which pain and/orinflammation predominates).

Compounds that are VR1 agonists may further be used, for example, incrowd control (as a substitute for tear gas) or personal protection(e.g., in a spray formulation) or as pharmaceutical agents for thetreatment of pain, itch, urinary incontinence or overactive bladder viacapsaicin receptor desensitization. In general, compounds for use incrowd control or personal protection are formulated and used accordingto conventional tear gas or pepper spray technology.

Within separate aspects, the present invention provides a variety ofnon-pharmaceutical in vitro and in vivo uses for the compounds providedherein. For example, such compounds may be labeled and used as probesfor the detection and localization of capsaicin receptor (in samplessuch as cell preparations or tissue sections, preparations or fractionsthereof). In addition, compounds provided herein that comprise asuitable reactive group (such as an aryl carbonyl, nitro or azide group)may be used in photoaffinity labeling studies of receptor binding sites.In addition, compounds provided herein may be used as positive controlsin assays for receptor activity, as standards for determining theability of a candidate agent to bind to capsaicin receptor, or asradiotracers for positron emission tomography (PET) imaging or forsingle photon emission computerized tomography (SPECT). Such methods canbe used to characterize capsaicin receptors in living subjects. Forexample, a VR1 modulator may be labeled using any of a variety of wellknown techniques (e.g., radiolabeled with a radionuclide such astritium, as described herein), and incubated with a sample for asuitable incubation time (e.g., determined by first assaying a timecourse of binding). Following incubation, unbound compound is removed(e.g., by washing), and bound compound detected using any methodsuitable for the label employed (e.g., autoradiography or scintillationcounting for radiolabeled compounds; spectroscopic methods may be usedto detect luminescent groups and fluorescent groups). As a control, amatched sample containing labeled compound and a greater (e.g., 10-foldgreater) amount of unlabeled compound may be processed in the samemanner. A greater amount of detectable label remaining in the testsample than in the control indicates the presence of capsaicin receptorin the sample. Detection assays, including receptor autoradiography(receptor mapping) of capsaicin receptor in cultured cells or tissuesamples may be performed as described by Kuhar in sections 8.1.1 to8.1.9 of Current Protocols in Pharmacology (1998) John Wiley & Sons, NewYork.

Compounds provided herein may also be used within a variety of wellknown cell separation methods. For example, modulators may be linked tothe interior surface of a tissue culture plate or other support, for useas affinity ligands for immobilizing and thereby isolating, capsaicinreceptors (e.g., isolating receptor-expressing cells) in vitro. Withinone preferred embodiment, a modulator linked to a fluorescent marker,such as fluorescein, is contacted with the cells, which are thenanalyzed (or isolated) by fluorescence activated cell sorting (FACS).

VR1 modulators provided herein may further be used within assays for theidentification of other agents that bind to capsaicin receptor. Ingeneral, such assays are standard competition binding assays, in whichbound, labeled VR1 modulator is displaced by a test compound. Briefly,such assays are performed by: (a) contacting capsaicin receptor with aradiolabeled VR1 modulator as described herein, under conditions thatpermit binding of the VR1 modulator to capsaicin receptor, therebygenerating bound, labeled VR1 modulator; (b) detecting a signal thatcorresponds to the amount of bound, labeled VR1 modulator in the absenceof test agent; (c) contacting the bound, labeled VR1 modulator with atest agent; (d) detecting a signal that corresponds to the amount ofbound labeled VR1 modulator in the presence of test agent; and (e)detecting a decrease in signal detected in step (d), as compared to thesignal detected in step (b).

The following Examples are offered by way of illustration and not by wayof limitation. Unless otherwise specified all reagents and solvent areof standard commercial grade and are used without further purification.Using routine modifications, the starting materials may be varied andadditional steps employed to produce other compounds provided herein.

EXAMPLES

Mass spectroscopy data in the following Examples is Electrospray MS,obtained in positive ion mode using a Micromass Time-of-Flight LCT(Micromass, Beverly, Mass.), equipped with a Waters 600 pump (WatersCorp., Milford, Mass.), Waters 996 photodiode array detector, Gilson 215autosampler (Gilson, Inc. Middleton, Wis.), and a Gilson 841microinjector. MassLynx (Advanced Chemistry Development, Inc; Toronto,Canada) version 4.0 software with OpenLynx processing was used for datacollection and analysis. MS conditions are as follows: capillaryvoltage=3.5 kV; cone voltage=30 V, desolvation and sourcetemperature=350° C. and 120° C., respectively; mass range=181-750 with ascan time of 0.22 seconds and an interscan delay of 0.05 min.

Sample volume of 1 microliter is injected onto a 50×4.6 mm ChromolithSpeedROD RP-18e column (Merck KGaA, Darmstadt, Germany), and elutedusing a 2-phase linear gradient at 6 ml/min flow rate. Sample isdetected using total absorbance count over the 220-340 nm UV range.Elution conditions are: Mobile Phase A—95/5/0.05 Water/MeOH/TFA; MobilePhase B—5/95/0.025 Water/MeOH/TFA. The following gradient is used(inject to inject cycle of 2.2 min):

Time(min) % B Gradient: 0 10 0.5 100 1.2 100 1.21 10

Example 1 Preparation of Representative Substituted BiarylPiperazinyl-Pyridine Analogues

This Example illustrates the preparation of representative substitutedbiaryl piperazinyl-pyridine analogues.

A.(5-Chloro-6-{4-[1-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyridin-3-yl)-aceticacid 1.(5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyridin-3-yl)-acetonitrile

To an ice cooled solution of(5-chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyridin-3-yl)-methanol(136 mg, 0.282 mmol) and Et₃N (79 μL, 0.564 mmol) in DCM, add MsCl (33μL, 0.422 mmol) dropwise. Remove ice bath and stir for 1 h at roomtemperature. Dilute with 3 volumes of DCM, wash with brine, dry theorganic layer (Na₂SO₄) and concentrate under reduced pressure. Dissolvethe crude mesylate in DMSO, add 138 mg of NaCN and heat at 60° C. untilno starting material remains as indicated by TLC. Dilute with water andextract the aqueous solution with EtOAc. Wash the organic extract withwater (2×) followed by brine (1×), dry (Na₂SO₄) and concentrate underreduced pressure to give crude product. Purify using flashchromatography to give pure title compound.

2.(5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyridin-3-yl)-aceticacid

Heat a solution of(5-chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyridin-3-yl)-acetonitrile(50 mg, 0.098 mmol) in 12 M HCl for 4 h. Concentrate the mixture underreduced pressure and then place under vacuum to remove remaining HCl.Add a small amount of water and extract the resulting white precipitatewith DCM. Dry the DCM (Na₂SO₄) and concentrate under reduced pressure.Triturate the resulting residue with ether to afford the title compoundas a pale yellow solid.

B.5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-nicotinicacid 1.4-(3-Chloro-5-ethoxycarbonyl-pyridin-2-yl)-piperazine-1-carboxylic acidtert-butyl ester

Heat a solution of 5,6-dichloro-nicotinic acid ethyl ester (TCI America)(4.43 g, 0.02 mol), piperazine-1-carboxylic acid tert-butyl ester (4.13g, 0.022 mol) and DIEA (5.2 mL, 0.03 mol) in DMA at 110° C. for 5 h.Partition the reaction mixture between water and EtOAc. Wash the EtOAclayer with water (1×) and brine (1×), dry (Na₂SO₄) and concentrate underreduced pressure to give the title compound as an orange oil.

2. 5-Chloro-6-piperazin-1-yl-nicotinic acid ethyl ester

Dissolve a solution of4-(3-chloro-5-ethoxycarbonyl-pyridin-2-yl)-piperazine-1-carboxylic acidtert-butyl ester (7.78 g, 0.021 mol) in dioxane, and then add a solutionof 4M HCl/dioxane (12 mL). Stir at 50° C. for 2 h, and then addadditional 4M HCl/dioxane (5 mL) and stir for an additional 1 h. Coolthe mixture in ice, collect the precipitate and wash with ether. Preparethe freebase by partitioning between EtOAc and 10% NaOH solution. Drythe organic layer (Na₂SO₄) and concentrate under reduced pressure togive the title compound.

3. 2,4-dichloro-6-(4-fluorophenyl)pyrimidine

Dissolve 4-fluorobromobenzene (8.75 g, 0.05 moles) in anhydrous ether(80 mL) under nitrogen atmosphere and cool to −78° C. Add dropwise 1.6 Mn-BuLi (34 mL, 0.055 moles) and stir at −78° C. for 45 min. Dissolve2,4-dichloropyrimidine (7.45 g, 0.05 moles) in Et₂O (100 mL) and adddropwise to the reaction mixture and warm the reaction mixture to −30°C. and stir at this temperature for 30 min followed by 0° C. for 30 min.Quench the reaction mixture with HOAc (3.15 mL, 0.055 moles) and water(0.5 mL, 0.027 moles) dissolved in THF (5.0 mL). Add dropwise a THF (40mL) solution of DDQ (11.9 g, 0.053 moles) to the reaction mixture. Bringthe reaction mixture to room temperature and stir at room temperaturefor 30 min. Cool the reaction mixture to 0° C. and add 3.0 N aq. NaOH(35 mL) and stir for 30 min. Decant the organic layer from the reactionmixture and wash the brown solid with Et₂O (3×100 mL). Combine theorganic layers, wash several times with saturated NaCl solution and drywith MgSO₄. Filter and evaporate under vacuum to afford brown coloredsolid. Purify the crude by flash column chromatography using 5%EtOAc/hexane to afford the title product as white solid.

4.5-Chloro-6-{4-[2-chloro-6-(4-fluoro-phenyl)-pyrimidin-4-yl]piperazin-1-yl}-nicotinicacid ethyl ester

To a mixture of 2,4-dichloro-6-(4-fluoro-phenyl)-pyrimidine (2.82 g,0.12 mol) and NaHCO₃ (1.95 g, 0.023 mol) in EtOH at 0° C., add5-chloro-6-piperazin-1-yl-nicotinic acid ethyl ester (3.44 g, 0.013mol). Remove the ice bath and stir at room temperature for 16 h.Concentrate under reduced pressure and partition between EtOAc andbrine. Dry the organic layer (Na₂SO₄) and concentrate under reducedpressure to give the title compound.

5.5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-nicotinicacid ethyl ester

Heat a mixture of5-chloro-6-{4-[2-chloro-6-(4-fluoro-phenyl)-pyrimidin-4-yl]-piperazin-1-yl}-nicotinicacid ethyl ester (1.19 g, 0.0025 mol) and 2-methylpyrrolidine (2.5 mL,0.025 mol) in DMA at 120° C. for 14 h. Partition the reaction mixturebetween EtOAc and water. Wash the organic layer with water (1×) andbrine (1×), then dry (Na₂SO₄) and concentrate under reduced pressure.Filter the crude product through a silica gel pad (2 inches deep×3inches in diameter) eluting with 700 mL of 30% EtOAc/hexanes.Concentrate under reduced pressure to give the title compound as anoff-white foam.

6.5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-nicotinicacid

To a solution of5-chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-nicotinicacid ethyl ester (427 mg, 0.813 mmol) in THF, add water dropwise untilthe cloudiness almost persists. To this mixture add LiOH.H₂O (350 mg,8.13 mmol) followed by a small amount of EtOH. Heat the mixture at 55°C. for 2 h, and then concentrate under reduced pressure. Add a smallamount of water to the residue, followed by 8.13 mmol of HCl (3Msolution). Adjust the final pH to 4 and collect the off-white solid viafiltration. Wash the solid with water and dry to afford the titlecompound.

C.(5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyridin-3-ylmethyl)-dimethyl-amine 1.(5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyridin-3-yl)-methanol

Dissolve5-chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-nicotinicacid ethyl ester (595 mg, 1.13 mmol, prepared in Example 1-B, step 5,above) in DCM and cool to −78° C. using a dry ice/acetone bath. Adddiisobutylaluminum hydride (4.53 mL, 1N in hexanes) dropwise andcontinue stirring for 1 h at −78° C. Add excess Na₂SO₄.10H₂O, stir at−78° C. for 5 min and then remove the cooling bath and allow to come toroom temperature. Filter the mixture through celite washing with DCMthen concentrate under reduced pressure. The resulting oil is purifiedusing flash chromatography (30-50% EtOAc/hexanes eluent) to afford thetitle compound as a white foam.

2. Methanesulfonic acid5-chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyridin-3-ylmethylester

To an ice cooled solution of(5-chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyridin-3-yl)-methanol(136 mg, 0.282 mmol) and Et₃N (79 μL, 0.564 mmol) in DCM, add MsCl (33μL, 0.422 mmol) dropwise. Remove ice bath and stir for 1 h at roomtemperature. Dilute with 3 volumes of DCM, wash with brine, dry theorganic layer (Na₂SO₄) and concentrate under reduced pressure to givethe title mesylate.

3.(5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyridin-3-ylmethyl)-dimethyl-amine

Heat a mixture of methanesulfonic acid5-chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyridin-3-ylmethylester with an excess of dimethylamine (1M in THF) in a sealed tube at80° C. for 2 h. Concentrate under reduced pressure then partitionbetween EtOAc and 10% NaOH solution. Dry the organic layer (Na₂SO₄) andconcentrate under reduced pressure. Purify the resulting residue usingflash chromatography (10:90:1/MeOH:DCM:NH₄OH eluent) to give the titlecompound.

D.6-{4-[6-(4-Fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinicacid 1. 4,6-Dichloro-2-(2-methylpyrrolidin-1-yl)pyrimidine

Dissolve 2,4,6-trichloropyrimidine (23.5 g, 0.13 mol) in anhydrous MeOH(220 mL), and add solid sodium bicarbonate (28.3 g, 0.33 mol). Cool to0° C. and add 2-methylpyrrolidine (12 g, 0.14 mol) dropwise. Let stir atroom temperature for 16 h. Filter off the excess sodium bicarbonate, andevaporate under reduced pressure. Purify by SiO₂ flash chromatography,using 50:1 hexanes:EtOAc, increasing to 30:1 hexanes:EtOAc, to providethe title compound as a white solid.

2. 4-Chloro-6-(4-fluorophenyl)-2-(2-methylpyrrolidin-1-yl)pyrimidine

Heat a mixture of 4,6-dichloro-2-(2-methylpyrrolidin-1-yl)pyrimidine(2.25 g, 9.74 mmol), 4-fluorophenylboronic acid (10.23 mmol), Pd(PPh₃)₄(562 mg, 0.487 mmol), and a 2M potassium phosphate solution (9.74 mL) indioxane (35 mL) under nitrogen at 80° C. for 16 h. Evaporate the mixtureand add water (50 mL). Extract with EtOAc (3×50 mL), dry (Na₂SO₄), andevaporate. Purify by SiO₂ flash chromatography, eluting with 9:1hexanes:EtOAc to provide the title compound as an oil.

3. 1-(5-Bromo-3-methyl-pyridin-2-yl)-3-(R)-methyl-piperazine

Heat a solution of 2,5-dibromo-3-methyl-pyridine (Chontech Inc.) (2.0 g,7.97 mmol), (R)-2-methyl-piperazine (3.2 g, 31.9 mmol) in DMA at 130° C.for 16 h. Partition the reaction mixture between water and EtOAc. Washthe EtOAc layer with water (1×) and brine (1×), dry (Na₂SO₄) andconcentrate under reduced pressure to give the title compound as asolid.

4.4-[4-(5-Bromo-3-methyl-pyridin-2-yl)-2-(R)-methyl-piperazin-1-yl]-6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidine

Heat a mixture of4-chloro-6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidine(1.30 g, 4.44 mmol),1-(5-bromo-3-methyl-pyridin-2-yl)-3-(R)-methyl-piperazine (1.2 g, 4.44mmol), and NaHCO₃ (0.71 g, 8.46 mmol) in EtOH at 50° C. for 20 h.Concentrate under reduced pressure and partition between EtOAc andbrine. Dry the organic layer (Na₂SO₄) and concentrate under reducedpressure. Purify the residue by flash column chromatography eluting withEtOAc-hexanes (1:4) to afford the title compound as a white solid.

5.6-{4-[6-(4-Fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinonitrile

To a mixture of4-[4-(5-bromo-3-methyl-pyridin-2-yl)-2-(R)-methyl-piperazin-1-yl]-6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidine(700 mg, 1.33 mmol) and Zn(CN)₂ (94 mg, 0.799 mmol) in DMF, addPd(PPh₃)₄ (77 mg, 0.067 mmol). Purge the reaction mixture for 10 minwith dry N₂. Heat the stirring reaction mixture overnight at 80° C.,cool to room temperature and partition between water and EtOAc. Dry thesolution (Na₂SO₄), concentrate under reduced pressure. Purify theresidue by flash column eluting with EtOAc-Hexanes (1:1) to afford thetitle compound as a white solid.

6.6-{4-[6-(4-Fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinicacid

Heat a solution of6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinonitrile(100 mg, 0.212 mmol) in 12 M HCl for 4 h at 90° C. Concentrate themixture under reduced pressure. Add a small amount of water, adjust thepH to 6-7, and collect the resulting white precipitate to afford thetitle compound as an off-white solid.

E.6-{4-[6-(4-Fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinamide

To a solution of6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinicacid (70 mg, 0.142 mmol) in DCM, add oxalyl chloride (3 equivalents) and1 drop of DMF. Stir the solution for 1 h at room temperature,concentrate, and dissolve in DCM. Cool the solution in an ice-bath, passNH₃ through the solution for 15 min, and stir for 2 h at roomtemperature. Wash with water. Dry the solution (Na₂SO₄) and concentrateunder reduced pressure. Purify the residue by flash columnchromatography eluting with DCM-MeOH (9:1) to afford the title compoundas an off white solid. ¹H NMR (400 MHz, CDCl₃): δ 1.32 (m, 3H, CH(CH₃));1.70 (m, 1H, CH₂CH₂); 1.91 (m, 1H, CH₂CH₂); 2.05 (m, 2H, CH₂CH₂); 2.39(s, 3H, Ar—CH₃); 3.05 (m, 1H); 3.19 (m, 1H); 3.38 (m, 1H); 3.68 (m, 4H);4.35 (m, 2H); 4.70 (m, 1H); 5.84 (br, 2H, NH₂); 6.24 (s, 1H, Ar—H); 7.10(m, 2H); 7.90 (d, 1H, J=2.0 Hz); 8.01 (m, 2H); 8.52 (s, 1H).

F.(6-{4-[6-(4-Fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridin-3-yl)-methanol 1.6-{4-[6-(4-Fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinicacid methyl ester

Reflux a solution of6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinicacid (120 mg, 0.245 mmol) and 2 drops of concentrated H₂SO₄ in MeOH for4 h. Cool to room temperature, concentrate, and partition betweensaturated NaHCO₃ and EtOAc. Dry the solution (Na₂SO₄), concentrate underreduced pressure. Purify the residue by flash column chromatographyeluting with EtOAc-Hexanes (1:4) to afford the title ester as an oil.

2.(6-{4-[6-(4-Fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridin-3-yl)-methanol

Dissolve6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinicacid methyl ester (78 mg, 0.155 mmol) in DCM and cool to −78° C. using adry ice/acetone bath. Add diisobutylaluminum hydride (0.619 mL, 1M inhexanes) dropwise and continue stirring for 1 h at −78° C. Add excessNa₂SO₄.10H₂O and stir at −78° C. for 5 min and then remove the coolingbath and allow to come to room temperature. Filter the mixture throughcelite washing with DCM, and then concentrate under reduced pressure.Purify the resulting oil using flash chromatography (30-50%EtOAc/hexanes eluent) to afford the title compound as a white foam. ¹HNMR (400 MHz, CDCl₃): δ 1.32 (m, 3H, CH₃); 1.40 (m, 3H, CH₃); 1.69 (m,1H, CH₂CH₂); 1.90 (m, 1H, CH₂CH₂); 2.05 (m, 2H, CH₂CH₂); 2.36 (s, 3H,Ar—CH₃); 3.00 (m, 1H); 3.08 (m, 1H); 3.37 (m, 2H); 3.48 (m, 1H); 3.68(m, 2H); 4.34 (m, 2H); 4.69 (s, 2H, CH₂OH); 4.71 (m, 1H); 6.24 (s, 1H,Ar—H); 7.10 (m, 2H); 7.48 (d, 1H, J=2.0 Hz); 8.00 (m, 2H); 8.11 (s, 1H).

G.6-{4-[6-(4-Fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-methyl-nicotinamidine

Pass HCl gas through a cooled solution of6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-methyl-nicotinonitrile(100 mg, 0.219 mmol) in EtOH (30 ml) for 15 min. Keep the solution at 5°C. for 24 h and concentrate under reduce pressure. Add 30 ml of EtOH,cool to 0° C., and pass NH₃ gas through the solution for 20 min. Keepthe reaction mixture at room temperature for 48 h. Concentrate underreduce pressure and purify the residue by silica gel plug eluting withDCM:MeOH:NH₄OH (90:10:1) to afford the title compound as off-whitesolid. ¹H NMR (300 MHz, CDCl₃): δ 1.19-1.25 (m, 3H, CH(CH₃)); 1.58-1.62(m, 1H, CH₂CH₂); 1.75-1.84 (m, 1H, CH₂CH₂); 1.93-2.01 (m, 2H, CH₂CH₂);2.22 (s, 3H, Ar—CH₃); 3.26 (br, 4H, pip-H); 3.58 (br, 5H); 4.24 (br,1H); 5.70 (m, 2H); 6.15 (s, 1H, Ar—H); 7.95 (m, 2H); 8.10 (s, 1H); 8.72(s, 1H); 8.98 (br, 3H, C(═NH)NH₂).

Example 2 Synthesis of Additional Representative Substituted BiarylPiperazinyl-Pyridine Analogues A.[2-(6-{4-[6-(3-Chloro-4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-methyl-pyridin-3-yloxy)-ethyl]-dimethyl-amine 1.1-(3-Methyl-5-nitro-pyridin-2-yl)-piperazine

1-(3-Methyl-5-nitro-pyridin-2-yl)-piperazine is prepared by heating asolution of 1 g of 2-chloro-3-methyl-5-nitro-pyridine (MaybridgeChemical Company Ltd.) and piperazine (5 g) in DMA at 110° C. for 16 h.Partition between EtOAc and water and wash the organic layer with water(2×). Dry the organic layer with Na₂SO₄ and concentrate to give crudeproduct. Purify via flash chromatography eluting with MeOH:DCM:NH₄OH(9:90:1) to give the title compound.

2.4-(3-Chloro-4-fluoro-phenyl)-6-[4-(3-methyl-5-nitro-pyridin-2-yl)-piperazin-1-yl]-2-(2-methyl-pyrrolidin-1-yl)-pyrimidine

Heat a mixture of4-chloro-6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidine(210 mg, 0.718 mmol),1-(5-bromo-3-methyl-pyridin-2-yl)-3-(R)-methyl-piperazine (1.2 g, 4.44mmol), and DIEA (185 mg, 1.44 mmol) in DMA at 120° C. for 16 h.Concentrate under reduced pressure and partition between EtOAc andbrine. Dry the organic layer (Na₂SO₄) and concentrate under reducedpressure. Purify the residue by flash column eluting with EtOAc-Hexanes(1:4) to afford the title compound as a yellow solid.

3.6-{4-[6-(3-Chloro-4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-methyl-pyridin-3-ylamine

Heat a mixture of4-chloro-6-(3-chloro-4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidine(210 mg), 1-(3-Methyl-5-nitro-pyridin-2-yl)-piperazine, and DIEA (185mg, 1.44 mmol) in DMA at 120° C. for 16 h. Cool to room temperature andpartition between EtOAc and 1N NaOH. Wash with water, dry the solution(Na₂SO₄), and concentrate under reduced pressure. Purify the residue byflash column eluting with EtOAc to afford the title compound as alight-yellow solid. ¹H NMR (400 MHz, CDCl₃): δ 1.30 (t, 3H, J=6.4 Hz,CH₃); 1.69 (m, 1H, CH₂CH₂); 1.91 (m, 1H, CH₂CH₂); 2.06 (m, 2H, CH₂CH₂);2.28 (s, 3H, Ar—CH₃); 3.10 (m, 4H); 3.47 (br, 2H, NH₂); 3.68 (m, 4H);3.77 (m, 4H); 4.34 (m, 1H); 6.25 (s, 1H, Ar—H); 6.88 (d, J=2.0 Hz, 1H);7.18 (m, 1H); 7.70 (d, 1H, J=3.2 Hz); 7.89 (m, 1H); 8.07 (m, 1H).

4.6-{4-[6-(3-Chloro-4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-methyl-pyridin-3-ol

To a cooled solution (0° C.) of6-{4-[6-(3-chloro-4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-methyl-pyridin-3-ylamine(146 mg, 0.30 mmol) in 10% H₂SO₄, add a solution of NaNO₂ (22 mg, 0.32mmol) in 3 mL H₂O and stir at 0° C. for 30 min. Warm up to roomtemperature and heat at 90° C. for 1 h. Cool to room temperature, adjustpH to 7, and extract with EtOAc. Wash with brine, dry the solution(Na₂SO₄), and concentrate under reduced pressure. Purify the residue byflash column chromatography eluting with EtOAc:hexanes (1:1) to affordthe title compound.

5.[2-(6-{4-[6-(3-Chloro-4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-methyl-pyridin-3-yloxy)-ethyl]-dimethyl-amine

To a cooled mixture of (2-chloro-ethyl)-dimethyl-amine hydrochloride(116 mg, 0.81 mmol) in DMF, add Cs₂CO₃ (526 mg, 1.62 mmol) and stir at0° C. for 30 min. Add6-{4-[6-(3-chloro-4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-methyl-pyridin-3-ol(78 mg, 0.16 mmol) and NaI (29 mg, 0.16 mmol). Stir the mixture at roomtemperature for 1 h and then for 2 h at 45° C. Partition between EtOAcand water. Wash with brine, dry the solution (Na₂SO₄), and concentrateunder reduced pressure. Purify the residue by flash columnchromatography eluting with DCM:MeOH:NH₄OH (90:10:1) to afford the titlecompound. ¹H NMR (300 MHz, CDCl₃): δ1.29 (t, 3H, J=6.3 Hz, CH₃); 1.69(m, 1H, CH₂CH₂); 1.91 (m, 1H, CH₂CH₂); 2.05 (m, 2H, CH₂CH₂); 2.32 (s,3H, Ar—CH₃); 2.33 (s, 6H, N(CH₃)₂)_(;) 2.71 (t, J=5.7 Hz, 2H); 3.13 (m,4H); 3.64 (m, 4H); 3.80 (m, 4H); 4.06 (t, J=5.7 Hz, 2H); 4.34 (m, 1H);6.25 (s, 1H, Ar—H); 7.10 (d, J=2.7 Hz, 1H); 7.18 (m, 1H); 7.89 (m, 2H);8.04 (m, 1H).

B.5-Methyl-6-{4-[2-methylpyrrolidin-1-yl)-6-pyridin-4-yl]-piperazin-1-yl}-nicotinicacid ethyl ester 1. 4,6-Dichloro-2-(2-methylpyrrolidin-1-yl)pyrimidine

Dissolve 2,4,6-trichloropyrimidine (23.5 g, 0.13 mol) in anhydrous MeOH(220 mL), and add solid sodium bicarbonate (28.3 g, 0.33 mol). Cool to0° C. and add 2-methylpyrrolidine (12 g, 0.14 mol) dropwise. Let stir atroom temperature for 16 h. Filter off the excess sodium bicarbonate, andevaporate under reduced pressure. Purify by SiO₂ liquid chromatography,using 50:1 hexanes:EtOAc, increasing to 30:1 hexanes:EtOAc, to providethe title compound as a white solid.

2. 4-(5-bromo-3-methyl-pyridin-2-yl)-piperazine-1-carboxylic acidtert-butyl ester

Heat a solution of 2,5-dibromo-3-methyl-pyridine (Chontech Inc.) (12.6g, 50.3 mmol), piperazine-1-carboxylic acid tert-butyl ester (11.7 g,62.9 mmol) and DIEA (13.3 g, 102.0 mmol) in DMA at 130° C. for 36 h.Partition the dark brown reaction mixture between water and EtOAc. Washthe EtOAc layer with water (1×) and brine (1×), dry (Na₂SO₄) andconcentrate under reduced pressure to give the crude product. Purify byflash chromatography, eluting with 9:1 hexanes:EtOAc, increasing to 3:1hexanes:EtOAc to provide the title compound as a solid.

3. 4-(5-Cyano-3-methyl-pyridin-2-yl)-piperazine-1-carboxylic acidtert-butyl ester

Heat a solution of4-(5-bromo-3-methyl-pyridin-2-yl)-piperazine-1-carboxylic acidtert-butyl ester (5.0 g, 14.0 mmol), Zn(CN)₂ (989 mg, 8.4 mmol) andPd(PPh₃)₄ (970 mg, 0.84 mmol), in dry DMF (50 mL) at 80° C. for 16 hunder nitrogen in a sealed tube. Partition the reaction mixture betweenwater and EtOAc. Wash the EtOAc layer with water (1×) and brine (1×),dry (Na₂SO₄) and concentrate under reduced pressure to give the titlecompound, and use immediately for the next reaction.

4. 5-Methyl-6-piperazine-1-yl-nicotinic acid

Treat a mixture of4-(5-cyano-3-methyl-pyridin-2-yl)-piperazine-1-carboxylic acidtert-butyl ester (5 g) with concentrated HCl (75 mL). Let stir at roomtemperature for 5 min or until the outgassing has ceased, then heat in asealed tube at 90° C. for 4 h. Evaporate the mixture, triturate withether, and collect the solid 5-methyl-6-piperazine-1-yl-nicotinic acid.

5. 5-Methyl-6-piperazine-1-yl-nicotinic acid ethyl ester

Place the carboxylic acid (5 g) in dry EtOH (100 mL) and bubble in HCl(g) for 10 min, then heat at 60° C. in a sealed tube for 16 h. Evaporatethe mixture, add 1M NaOH (100 mL), and extract with EtOAc (3×50 mL). Dry(Na₂SO₄) and evaporate to furnish the title compound as a tan oil.

6.6-{4-[6-chloro-2-(2-methylpyrrolidin-1-yl]-piperazin-1-yl}-5-methyl-nicotinicacid ethyl ester

Heat a mixture of 5-methyl-6-piperazine-1-yl-nicotinic acid ethyl ester(2.0 g, 8.0 mmol), 4,6-dichloro-2-(2-methyl-pyrrolidin-1-yl)pyrimidine(1.9 g, 8.2 mmol), and sodium bicarbonate (1.4 g, 16.0 mmol) in EtOH (50mL) for 16 h. Evaporate the mixture and place directly on a silica gelcolumn. Elute with 3:1 hexanes:EtOAc to give the title compound as awhite foamy solid.

7.5-Methyl-6-{4-[2-methylpyrrolidin-1-yl)-6-pyridin-4-yl-pyrimidin-4-yl]-piperazin-1-yl}-nicotinicacid ethyl ester

Heat a mixture of6-{4-[6-chloro-2-(2-methylpyrrolidin-1-yl]-piperazin-1-yl}-5-methyl-nicotinicacid ethyl ester (300 mg, 0.7 mmol), 4-tri-n-butylstannylpyridine (526mg, 1.4 mmol) and Pd(PPh₃)₄ (39 mg, 0.03 mol) in toluene (15 mL) at 110°C. for 16 h. Let cool to room temperature, filter off the catalyst, andadd water (10 mL). Extract with EtOAc (3×10 mL), dry (Na₂SO₄) andevaporate. Purify using flash chromatography (9:1 hexanes/EtOAc) to givepure title compound. ¹H NMR (CDCl₃): 8.78 (s, 1H), 8.70 (d, 2H), 8.00(s, 1H), 7.92 (d, 2H), 6.38 (s, 1H), 4.40 (quart, 2H), 3.81 (mult, 4H),3.75 (mult, 1H), 3.41 (mult, 4H), 2.40 (s, 3H), 2.10 (m, 2H), 1.62 (m,2H), 1.60 (m, 2H).

C.6-{4-[6-(2-Isopropyl-pyridin-4-yl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-methyl-nicotinicacid ethyl ester 1. 2-Isopropyl-4-trimethylstannanyl-pyridine

To a cold suspension (0° C.) of Na (25% in toluene, 10 g, 104 mmol) inDME (100 mL) add a solution of trimethyltin chloride (9.4 g, 47.4 mmol)in DME (20 mL) dropwise. Stir the mixture at 0° C. for 3 h followed byaddition of the solution of cold (0° C.) 4-chloro-2-isopropyl-pyridine(Comins & Mantlo (1985) J. Org. Chem. 50:4410-4411) (4.9 g, 31.6 mmol)in DME (20 mL). Stir the mixture at 0° C. for 2 h and warm to roomtemperature. Filter, concentrate the filtrate, and dilute the residuewith ether. Filter and concentrate the filtrate. Distil the residue (bp75-80° C. at 1 mm Hg) to afford the title compound as a light oil.

2.6-{4-[6-(2-Isopropyl-pyridin-4-yl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-methyl-nicotinicacid ethyl ester

Using a procedure analogous to that used in Example 2-B step 7,6-{4-[6-chloro-2-(2-methylpyrrolidin-1-yl]-piperazin-1-yl}-5-methyl-nicotinicacid ethyl ester is reacted with2-isopropyl-4-trimethylstannanyl-pyridine to give the title compound.

D.5-Chloro-6-{4-[2-(3-chloro-4-fluoro-phenyl)-6-morpholin-4-yl-pyridin-4-yl]-piperazin-1-yl}-nicotinicacid ethyl ester 1. 2-chloro-6-morpholino-4-yl-pyridin-4-ylamine

Stir a solution of 4-amino-2,6-dichloropyridine (3.3 g) in morpholine(15 mL) for 4 h at 150° C., concentrate, partition between H₂O andEtOAc, dry over Na₂SO₄, and concentrate under vacuum. Purify by flashchromatography (2:3 hexanes/EtOAc) to give the title compound.

2. 2-(3-chloro-4-fluoro-phenyl)-6-morpholino-4-yl-pyridin-4-ylamine

To a de-gassed mixture of 3-chloro-4-fluorophenylboronic acid (849 mg,4.87 mmol), 2-chloro-6-morpholino-4-yl-pyridin-4-ylamine (800 mg, 3.74mmol), and 2M K₃PO₄ (7.5 mmol), in dioxane (15 mL) under nitrogen addPd(PPh₃)₄ (0.23 mmol). Stir the mixture at 80° C. for 16 h, concentrate,extract with EtOAc. Dry over Na₂SO₄, concentrate under vacuum, andpurify by flash chromatography (1:1 hexanes/EtOAc) to give the titlecompound.

3. 4-{4-bromo-6-(3-chloro-4-fluoro-phenyl)-pyridin-2yl}-morpholine

To an ice cooled solution of2-(3-chloro-4-fluoro-phenyl)-6-morpholino-4-yl-pyridin-4-ylamine (250mg, 0.81 mmol) in 75% H₂SO₄ (10 mL) add dropwise a solution of NaNO₂ (56mg, 0.81 mmol) in 3 mL H₂O. Stir the mixture 30 min at 0° C. Add CuBr(135 mg, 0.93 mmol) and 48% HBr (2 mL). Stir the mixture 15 min at 0° C.then 30 min at 60° C. Cool to room temperature, neutralize to pH 8,extract with EtOAc, dry over Na₂SO₄, and concentrate under vacuum.Purify by flash chromatography (3:1 hexanes/EtOAc) to give the titlecompound.

4.5-Chloro-6-{4-[2-(3-chloro-4-fluoro-phenyl)-6-morpholin-4-yl-pyridin-4-yl]-piperazin-1-yl}-nicotinicacid ethyl ester

To a de-gassed mixture of4-{4-bromo-6-(3-chloro-4-fluoro-phenyl)-pyridin-2yl}-morpholine (50 mg,0.135 mmol), 5-chloro-6-piperazin-1-yl-nicotinic acid ethyl ester (0.162mmol), and 1M (THF) t-BuOK (0.162 mmol), in toluene (3 mL) undernitrogen add Pd₂(dba)₃ (0.0054 mmol) and BINAP (0.0067 mmol). Stir themixture at 80° C. for 16 h, concentrate, extract with EtOAc. Dry overNa₂SO₄, concentrate under vacuum, and purify by preparative TLC (3:1hexanes/EtOAc) to give the title compound.

E.5-Chloro-6-{4-[4-(3-chloro-4-fluoro-phenyl)-6-morpholin-4-yl-pyridin-2-yl]-piperazin-1-yl}-nicotinicacid ethyl esteroline 1.2,3-dichloro-4-(3-chloro-4-fluoro-phenyl)-pyridine

To a de-gassed mixture of 3-chloro-4-fluorophenylboronic acid (77 mg,0.44 mmol)), 4-bromo-2,6-dichloro-pyridine (Talik and Plazek (1959)Rocz. Chem. 33:387-392) (100 mg, 0.44 mmol), and 2M Na₂CO₃ (0.55 mmol),in DME (4 mL) under nitrogen add Pd(PPh₃)₄ (0.026 mmol). Stir themixture at 80° C. for 16 h, concentrate, extract with EtOAc. Dry overNa₂SO₄, concentrate under vacuum, and purify by preparative TLC (9:1hexanes/EtOAc) to give the title compound.

2. 4-[6-chloro-4-(3-chloro-4-fluoro-phenyl)-pyridin-2-yl]-morpholine

Stir a solution of 2,6-dichloro-4-(3-chloro-4-fluoro-phenyl)-pyridine(100 mg) in morpholine (2 mL) 3 h at 80° C., concentrate, partitionbetween H₂O and EtOAc, dry over Na₂SO₄, and concentrate under vacuum.Purify by preparative TLC (3:1 hexanes/EtOAc) to give the titlecompound.

3.5-Chloro-6-{4-[4-(3-chloro-4-fluoro-phenyl)-6-morpholin-4-yl-pyridin-2-yl]-piperazin-1-yl}-nicotinicacid ethyl ester

To a de-gassed mixture of4-[6-chloro-4-(3-chloro-4-fluoro-phenyl)-pyridin-2-yl]-morpholine (50mg, 0.153 mmol)), 5-chloro-6-piperazin-1-yl-nicotinic acid ethyl ester(0.183 mmol), and 1M (THF) t-BuOK (0.183 mmol), in toluene (3 mL) undernitrogen add Pd₂(dba)₃ (0.006 mmol) and BINAP (0.008 mmol). Stir themixture at 80° C. for 16 h, concentrate, and extract with EtOAc. Dryover Na₂SO₄, concentrate under vacuum, and purify by preparative TLC(3:1 hexanes/EtOAc) to give the title ester.

F.(6-{4-[6-(4-Fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridin-3-yl)-aceticacid 1.4-[2-Chloro-6-(4-fluoro-phenyl)-pyrimidin-4-yl]-3-(R)-methyl-piperazine-1-carboxylicacid tert-butyl ester

Heat a mixture of 2,4-dichloro-6-(4-fluoro-phenyl)-pyrimidine (2.8 g,11.52 mmol), 3-(R)-methyl-piperazine-1-carboxylic acid tert-butyl ester(2.42 g, 12.1 mmol), and K₂CO₃ (3.2 g, 23.0 mmol) in DMA at 60° C. for16 h. Dilute with water, extract with EtOAc, and wash with brine. Drythe organic layer (Na₂SO₄) and concentrate under reduced pressure.Purify the residue by flash column eluting with EtOAc-Hexanes (1:4) toafford the title compound as a white solid.

2.4-[6-(4-Fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazine-1-carboxylicacid tert-butyl ester

Heat a mixture of4-[2-chloro-6-(4-fluoro-phenyl)-pyrimidin-4-yl]-3-(R)-methyl-piperazine-1-carboxylicacid tert-butyl ester (5.0 g, 12.3 mmol), 2-(R)-methyl-pyrrolidinehydrobromide (Nijhuis et. al. (1989) J. Org. Chem. 54:216-220) (3.0 g,16.0 mmol), and K₂CO₃ (5.2 g, 37.8 mmol) in DMA at 120° C. for 16 h.Dilute with water, extract with EtOAc, and wash with brine. Dry theorganic layer (Na₂SO₄) and concentrate under reduced pressure. Purifythe residue by flash column eluting with EtOAc-Hexanes (1:4) to affordthe title compound as a white solid.

3.4-(4-Fluoro-phenyl)-6-(2-(R)-methyl-piperazin-1-yl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidine

Stir4-[6-(4-fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazine-1-carboxylicacid tert-butyl ester (4.5 g, 9.88 mmol) in 4M HCl-dioxane (50 mL) for40 min. Concentrate and partition between EtOAc and sat. NaHCO₃. Dry theorganic layer (Na₂SO₄) and concentrate under reduced pressure to yieldthe title compound.

4.4-[4-(5-bromo-3-methyl-pyridin-2-yl)-2-(R)-methyl-piperazin-1-yl]-6-(4-fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidine

Heat a mixture of4-(4-fluoro-phenyl)-6-(2-(R)-methyl-piperazin-1-yl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidine(400 mg, 1.13 mmol), 2,5-dibromo-3-methyl-pyridine (367 mg, 1.46 mmol),and DIEA (218 mg, 1.69 mmol) in DMA at 135° C. for 96 h. Dilute withwater, extract with EtOAc, and wash with brine. Dry the organic layer(Na₂SO₄) and concentrate under reduced pressure. Purify the residue byflash column chromatography eluting with EtOAc-Hexanes (1:10) to affordthe title compound as a white solid.

5.(6-{4-[6-(4-Fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridin-3-yl)-aceticacid tert-butyl ester

To lithium dicyclohexyl amide add a solution of acetic acid tert-butylester (62 μL, 0.46 mmol) in toluene (2 mL). Stir for 10 min at roomtemperature, and then add4-[4-(5-bromo-3-methyl-pyridin-2-yl)-2-(R)-methyl-piperazin-1-yl]-6-(4-fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidine(200 mg, 0.38 mmol), Pd₂(dba)₃ (3.5 mg, 1%) and P(t-bu)₃ (0.8 mg, 1%).De-gas the suspension for 5 min and stir for 16 h at room temperature.Dilute with EtOAc, wash with brine, dry the organic layer (Na₂SO₄) andconcentrate under reduced pressure. Purify using flash chromatographyeluting with EtOAc-Hexanes (1:4) to give the title ester.

6.(6-{4-[6-(4-Fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridin-3-yl)-aceticacid

Stir a solution of(6-{4-[6-(4-fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridin-3-yl)-aceticacid tert-butyl ester (210 mg, 0.375 mmol) and 1 mL of TFA in DCM (10mL) at room temperature for 16 h. Concentrate and partition betweenEtOAc and sat. NaHCO₃. Dry the organic layer (Na₂SO₄) and concentrateunder reduced pressure. Purify using flash chromatography eluting withMeOH-DCM (1:19) to give the title compound.

G.6-{4-[6-(4-Fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridine-2-carboxylicacid 1.6-{4-[6-(4-Fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridine-2-carboxylicacid methyl ester

To a cold solution (0° C.) of 6-hydroxy-5-methyl-pyridine-2-carboxylicacid methyl ester (Adamczyk et. al. (2002) Tetrahedron 58:6951-6963)(167 mg, 1.0 mmol) in CHCl₃ (15 mL), add trifluoroacetic anhydride (423mg, 1.5 mmol) dropwise followed by TEA (202 mg, 2.0 mmol). Stir themixture for 1 h. Dilute with CHCl₃, wash with sat. NaHCO₃, dry theorganic layer (Na₂SO₄) and concentrate under reduced pressure. Mix theresidue with4-(4-fluoro-phenyl)-6-(2-(R)-methyl-piperazin-1-yl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidine(426 mg, 1.2 mmol) and DIEA (129 mg, 1.0 mmol) in DMA. Heat the mixtureat 100° C. for 16 h. Dilute with water, extract with EtOAc, and washwith brine. Dry the organic layer (Na₂SO₄) and concentrate under reducedpressure. Purify the residue by flash column eluting with EtOAc-Hexanes(1:5) to afford the title compound.

2.6-{4-[6-(4-Fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridine-2-carboxylicacid

To a solution of6-{4-[6-(4-fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridine-2-carboxylicacid methyl ester (300 mg, 0.595 mmol) in THF, add water dropwise untilthe cloudiness almost persists. To this mixture add LiOH.H₂O (50 mg, 1.2mmol). Heat the mixture at 50° C. for 2 h, and then concentrate underreduced pressure. Add a small amount of water to the residue. Adjust thefinal pH to 6 and collect the off-white solid via filtration. Wash thesolid with water and dry to afford the title compound.

H.6-{4-[6-(4-Fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridine-2-carboxylicacid amide

To a solution of6-{4-[6-(4-fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-pyridine-2-carboxylicacid (84 mg, 0.142 mmol) in DCM, add oxalyl chloride (3 equivalents) and1 drop of DMF. Stir the solution for 1 h at room temperature,concentrate, and dissolve in DCM. Cool the solution in an ice-bath, passNH₃ through the solution for 15 min, and stir for 2 h at roomtemperature. Wash with water. Dry the solution (Na₂SO₄) and concentrateunder reduced pressure. Purify the residue by flash columnchromatography eluting with DCM-MeOH (9:1) to afford the title compoundas an off white solid.

I.5-Chloro-6-{4-[6-(3-chloro-4-fluoro-phenyl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-nicotinicacid ethyl ester 1. 5-Chloro-6-(3-(R)-methyl-piperazin-1-yl)-nicotinicacid ethyl ester

Heat a solution of (R)-2-methyl-piperazine (0.07 mol),5,6-dichloro-nicotinic acid ethyl ester (TCI America) (10.1 g, 0.046mol) and potassium carbonate (31.7 g, 0.23 mol) in DMA at 110° C. for 48h. Cool the solution, and partition between EtOAc and brine. Separatethe layers and extract the aqueous portion with EtOAc (1×). Wash thecombined organic layers with 10% NaOH (4×), dry (Na₂SO₄), andconcentrate under reduced pressure to give an oil which crystallizes onstanding.

2.5-Chloro-6-[4-(6-chloro-pyrimidin-4-yl)-3(R)-methyl-piperazin-1-yl]-nicotinicacid ethyl ester

Heat a solution of 5-chloro-6-(3-(R)-methyl-piperazin-1-yl)-nicotinicacid ethyl ester (338 mg, 1.2 mmol), 4,6-dichloropyrimidine (179 mg, 1.2mmol) and potassium carbonate (328 mg, 2.4 mmol) in DMA at 80° C. for 16h. Partition between EtOAc and brine then separate layers and wash theorganic layer with 10% NaOH (3×) followed by brine. Dry (Na₂SO₄) andconcentrate under reduced pressure to give the title compound.

3.5-Chloro-6-{4-[6-(3-chloro-4-fluoro-phenyl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-nicotinicacid ethyl ester

Bubble nitrogen through a mixture of5-chloro-6-[4-(6-chloro-pyrimidin-4-yl)-3-(R)-methyl-piperazin-1-yl]-nicotinicacid ethyl ester (226 mg, 0.57 mmol), 3-chloro-4-fluorophenylboronicacid (99 mg, 0.57 mmol), K₃PO₄ (2M aqueous, 570 μL) and Pd(PPh₃)₄ (33mg, 0.03 mmol) in dioxane for 10 min. Heat the mixture to 80° C. for 16h under a nitrogen atmosphere. Cool the mixture and partition betweenEtOAc and water. Dry (Na₂SO₄) the organic layer and concentrate underreduced pressure to yield the crude product. Purify the residue withpreparative plate chromatography (2×2 mm thick) using 40% EtOAc/hexanesas the eluent to afford the title compound as a foam. ¹H NMR (400 MHz,CDCl₃): δ1.38 (m, 6H, 2×CH₃), 3.16 (m, 1H), 3.31 (dd, 1H, J=11 Hz), 3.51(m, 1H), 4.12 (d, 1H, J=12.8 Hz), 4.21 (d, 1H, J=13 Hz), 4.39 (m, 3H),4.77 (Br s, 1H), 6.82 (s, 1H), 7.25 (m, 1H), 7.89 (m, 1H), 8.07 (d, 1H,J=7 Hz), 8.18 (s, 1H), 8.71 (s, 1H), 8.79 (s, 1H).

J.1′-[6-(3-Chloro-4-fluoro-phenyl)-2-(4-propyl-piperazin-1-yl)-pyrimidin-4-yl]-3-methyl-5-nitro-1′,2′,3′,4′,5′,6′-hexahydro-[2,4′]bipyridinyl 1.4-[2-Chloro-6-(3-chloro-4-fluoro-phenyl)-pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester

To a mixture of 2,4-dichloro-6-(3-chloro-4-fluoro-phenyl)-pyrimidine(0.12 mol) and NaHCO₃ (1.95 g, 0.023 mol) in EtOH at 0° C., addpiperazine-1-carboxylic acid tert-butyl ester (0.013 mol). Remove theice bath and stir at room temperature for 16 h. Concentrate underreduced pressure and partition between EtOAc and brine. Dry the organiclayer (Na₂SO₄) and concentrate under reduced pressure to give the titlecompound.

2.4-[6-(3-Chloro-4-fluoro-phenyl)-2-(4-propyl-piperazin-1-yl)-pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester

Heat a solution of4-[2-chloro-6-(3-chloro-4-fluoro-phenyl)-pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester (2.5 g, 5.9 mmol), N-propylpiperazine (5.9 mmol)and DIEA (11.7 mmol) in DMA at 100° C. for 16 h. Cool, partition between10% NaOH and EtOAc and wash the organic layer with additional NaOHsolution (3×). Dry the organic layer (Na₂SO₄) and concentrate underreduced pressure to afford the title compound.

3.4-(3-Chloro-4-fluoro-phenyl)-6-piperazin-1-yl-2-(4-propyl-piperazin-1-yl)-pyrimidine

Dissolve4-[6-(3-chloro-4-fluoro-phenyl)-2-(4-propyl-piperazin-1-yl)-pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester (2.0 g) in dioxane and then add a 4M solution ofHCl in dioxane (15 mL). After the formation of precipitate stir thesuspension vigorously for 2 h. Collect the solid and wash with ether andthen freebase the material by partitioning between 10% NaOH solution andDCM. Separate the organic layer, dry (Na₂SO₄) and concentrate to givethe title compound as an off-white solid.

4.1′-[6-(3-Chloro-4-fluoro-phenyl)-2-(4-propyl-piperazin-1-yl)-pyrimidin-4-yl]-3-methyl-5-nitro-1′,2′,3′,4′,5′,6′-hexahydro-[2,4′]bipyridinyl

Heat a mixture of4-(3-chloro-4-fluoro-phenyl)-6-piperazin-1-yl-2-(4-propyl-piperazin-1-yl)-pyrimidine(300 mg, 0.72 mmol), 2-chloro-3-methyl-5-nitro-pyridine (149 mg, 0.86mmol) and DIEA (186 mg, 1.44 mmol) in DMA (4 mL) for 16 h at 110° C.Cool, partition between 10% NaOH and EtOAc and wash the organic layerwith additional NaOH solution (3×). Dry the organic layer (Na₂SO₄) andconcentrate under reduced pressure to afford the title compound.

K.3′-Chloro-4-[6-(4-fluoro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-methyl-3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-carboxylicacid 1.4-[4-(4-Fluoro-phenyl)-6-(2-methyl-piperazin-1-yl)-pyrimidin-2-yl]-3-methyl-morpholine

This compound is prepared using (R)-3-methylmorpholine (WO 02/064096) ina procedure analogous to that used for the preparation of4-(4-fluoro-phenyl)-6-(2-methyl-piperazin-1-yl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidinein Example 2F step 3.

2.6′-Amino-3′-chloro-4-[6-(4-fluoro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-methyl-3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-carboxylicacid methyl ester

Heat a mixture of4-[4-(4-fluoro-phenyl)-6-(2-methyl-piperazin-1-yl)-pyrimidin-2-yl]-3-(R)-methyl-morpholinewith 1.1 equivalents of 3-amino-5,6-dichloro-pyrazine-2-carboxylic acidmethyl ester (Cragoe et. al. J. Med. Chem., 1967, 10, 66-75) inisopropanol at 85° C. for 12 h. Concentrate the mixture under reducedpressure, and then partition between 10% NaOH and EtOAc. Dry the organiclayer (Na₂SO₄) and concentrate under reduced pressure to give the titlecompound.

3.6′-Bromo-3′-chloro-4-[6-(4-fluoro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-methyl-3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-carboxylicacid methyl ester

To a cooled (5° C.) and well stirred mixture of6′-amino-3′-chloro-4-[6-(4-fluoro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-methyl-3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-carboxylicacid methyl ester (10 g) in 48% HBr (75 mL) and glacial acetic acid (120mL), add a solution (16 mL) of bromine in acetic acid (6:1) over aperiod of 45 min. To this mixture add a solution of NaNO₂ (8.3 g) inwater (18 mL) while maintaining the temperature below 8° C. After theaddition is complete, stir the mixture for 30 min and then destroy theexcess bromine by adding 100 mL of 30% NaHSO₃. Neutralize the solutionto pH 8 by the dropwise addition of 20% NaOH and then extract withEtOAc. Wash the organic extracts with dilute NH₄OH, dry (Na₂SO₄) andconcentrate under reduced pressure to give the title compound.

4.3′-Chloro-4-[6-(4-fluoro-phenyl)-2-(3-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(R)-methyl-3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-carboxylicacid methyl ester

Stir a solution of6′-bromo-3′-chloro-4-[6-(4-fluoro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-methyl-3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-carboxylicacid methyl ester (3 g) in 100 mL of THF with 5% palladium on carbon(300 mg) under 1 atmosphere of hydrogen gas for several days. Filter themixture through celite and concentrate under reduced pressure. Partitionthe residue between saturated NaHCO₃ solution and EtOAc. Dry (Na₂SO₄)the organic layer and concentrate under reduced pressure. Purify usingflash column chromatography eluting with a gradient EtOAc/hexanessolvent system to give the title compound.

5.3′-Chloro-4-[6-(4-fluoro-phenyl)-2-(3-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(R)-methyl-3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-carboxylicacid

Stir a solution of3′-chloro-4-[6-(4-fluoro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-methyl-3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-carboxylicacid methyl ester (500 mg) in a mixture of 10% NaOH (6 mL) and EtOH (20mL) at room temperature for 16 h. Bring the mixture to pH 4 with 6 N HCland extract the solution with EtOAc. Dry the organic layer (Na₂SO₄) andconcentrate under reduced pressure to give the title compound.

L.2-{4-[6-(3-Chloro-4-fluorophenyl)-2-(2-methylpyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyrimidine-5-carboxylicacid amide 1.2-{4-[6-(3-Chloro-4-fluorophenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-bromopyrimidine

Heat a solution of 5-bromo-2-chloropyrimidine (Lancaster Chemicals, 565mg),4-(3-chloro-4-fluorophenyl)-2-(2-methylpyrrolidin-1-yl)-6-piperazin-1-yl-pyrimidine(1000 mg), DIEA (500 mg) and DMA (5 mL) at 110° C. for 3 h undernitrogen. Dilute with EtOAc (15 mL), wash with water (3×10 mL), dry(Na₂SO₄), and evaporate. Purify by silica gel chromatography, elutingwith 1:1 hexanes:EtOAc to provide the title compound.

2.2-{4-[6-(3-Chloro-4-fluorophenyl)-2-(2-methylpyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-pyrimidine-5-carbonitrile

Heat a mixture of2-{4-[6-(3-chloro-4-fluorophenyl)-2-(2-methylpyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}-5-bromopyrimidine(478 mg) and CuCN (270 mg) in DMF at 150° C. for 16 h in a round bottomflask that is open to the air. Let cool, dilute with EtOAc (15 mL), washwith water (3×10 mL), dry (Na₂SO₄), and evaporate. Purify by silica gelchromatography, eluting with 3:1 hexanes:EtOAc to provide the titlecompound.

M.6-{4-[6-(4-Fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-chloro-nicotinicacid 1. 4,6-Dichloro-2-(2-(S)-methylpyrrolidin-1-yl)pyrimidine

Dissolve 2,4,6-trichloropyrimidine (23.5 g, 0.13 mol) in anhydrous MeOH(220 mL), and add solid sodium bicarbonate (28.3 g, 0.33 mol). Cool to0° C. and add 2-(S)-methylpyrrolidine (12 g, 0.14 mol) dropwise. Letstir at room temperature for 16 h. Filter off the excess sodiumbicarbonate, and evaporate under reduced pressure. Purify by SiO₂ flashchromatography, using 50:1 hexanes:EtOAc, increasing to 30:1hexanes:EtOAc, to provide the title compound as a white solid.

2. 4-Chloro-6-(4-fluorophenyl)-2-(2-(S)-methylpyrrolidin-1-yl)pyrimidine

Heat a mixture of 4,6-dichloro-2-(2-(S)-methylpyrrolidin-1-yl)pyrimidine(2.25 g, 9.74 mmol), 4-fluorophenylboronic acid (10.23 mmol), Pd(PPh₃)₄(562 mg, 0.487 mmol), and a 2M potassium phosphate solution (9.74 mL) indioxane (35 mL) under nitrogen at 80° C. for 16 h. Evaporate the mixtureand add water (50 mL). Extract with EtOAc (3×50 mL), dry (Na₂SO₄), andevaporate. Purify by SiO₂ flash chromatography, eluting with 9:1hexanes:EtOAc to provide the title compound as an oil.

3. 1-(5-Bromo-3-chloro-pyridin-2-yl)-3-(R)-methyl-piperazine

Heat a solution of 2,5-dibromo-3-chloro-pyridine (Chontech Inc.) (2.0 g)and (R)-2-methyl-piperazine (3.2 g, 31.9 mmol) in DMA at 130° C. for 16h. Partition the reaction mixture between water and EtOAc. Wash theEtOAc layer with water (1×) and brine (1×), dry (Na₂SO₄) and concentrateunder reduced pressure to give the title compound as a solid.

4.4-[4-(5-Bromo-3-chloro-pyridin-2-yl)-2-(R)-methyl-piperazin-1-yl]-6-(4-fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidine

Heat a mixture of4-chloro-6-(4-fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidine(1.30 g, 4.44 mmol),1-(5-bromo-3-chloro-pyridin-2-yl)-3-(R)-methyl-piperazine (1.2 g), andNaHCO₃ (0.71 g, 8.46 mmol) in EtOH at 50° C. for 20 h. Concentrate underreduced pressure and partition between EtOAc and brine. Dry the organiclayer (Na₂SO₄) and concentrate under reduced pressure. Purify theresidue by flash column eluting with EtOAc-Hexanes (1:4) to afford thetitle compound as a white solid.

5.6-{4-[6-(4-Fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-chloro-nicotinonitrile

To a mixture of4-[4-(5-bromo-3-chloro-pyridin-2-yl)-2-(R)-methyl-piperazin-1-yl]-6-(4-fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidine(700 mg) and Zn(CN)₂ (94 mg) in DMF, add Pd(PPh₃)₄ (77 mg, 0.067 mmol).Purge the reaction mixture for 10 min with dry N₂. Heat the stirringreaction mixture overnight at 80° C., cool to room temperature andpartition between water and EtOAc. Dry the solution (Na₂SO₄),concentrate under reduced pressure. Purify the residue by flash columneluting with EtOAc-Hexanes (1:1) to afford the title compound as a whitesolid.

6.6-{4-[6-(4-Fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-chloro-nicotinicacid

Heat a solution of6-{4-[6-(4-fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-chloro-nicotinonitrile(100 mg) in 12 M HCl for 4 h at 90° C. Concentrate the mixture underreduced pressure. Add a small amount of water, adjust the pH to 6-7, andcollect the resulting white precipitate to afford the title compound asan off-white solid.

N.2-(5-chloro-6-((R)-4-(6-(3-chloro-4-fluorophenyl)-2-((R)-2-methylpyrrolidin-1-yl)pyrimidin-4-yl)-3-methylpiperazin-1-yl)pyridin-3-yl)-N,N-dimethylethanamine 1.2-(5-chloro-6-((R)-4-(6-(3-chloro-4-fluorophenyl)-2-((R)-2-methylpyrrolidin-1yl)pyrimidin-4-yl)-3-methylpiperazin-1-yl)pyridin-3-yl)acetonitrile

The title compound is prepared from(5-chloro-6-((R)-4-(6-(3-chloro-4-fluorophenyl)-2-((R)-2-methylpyrrolidin-1-yl)pyrimidin-4-yl)-3-methylpiperazin-1-yl)pyridin-3-yl)methanolusing a procedure analogous to that used in Example 1-A1.

2.2-(5-chloro-6-((R)-4-(6-(3-chloro-4-fluorophenyl)-2-((R)-2-methylpyrrolidin-1-yl)pyrimidin-4-yl)-3-methylpiperazin-1-yl)pyridin-3-yl)acetaldehyde

Add a solution of diisobutylaluminum hydride (1M in hexane, 333 μL) to astirring solution of2-(5-chloro-6-((R)-4-(6-(3-chloro-4-fluorophenyl)-2-((R)-2-methylpyrrolidin-1-yl)pyrimidin-4-yl)-3-methylpiperazin-1-yl)pyridin-3-yl)acetonitrile(120 mg, 0.22 mmol) in dichloromethane at −78° C. over a period of 10min. When starting material is consumed, add an excess of Na₂SO₄.10H₂Oand stir for 1 h while bringing the reaction mixture to ambienttemperature. Filter the mixture through celite washing with DCM.Concentrate under reduced pressure and use without further purification.

3.2-(5-chloro-6-((R)-4-(6-(3-chloro-4-fluorophenyl)-2-((R)-2-methylpyrrolidin-1-yl)pyrimidin-4-yl)-3-methylpiperazin-1-yl)pyridin-3-yl)-N,N-dimethylethanamine

To a solution of2-(5-chloro-6-((R)-4-(6-(3-chloro-4-fluorophenyl)-2-((R)-2-methylpyrrolidin-1-yl)pyrimidin-4-yl)-3-methylpiperazin-1-yl)pyridin-3-yl)acetaldehyde(64 mg, 0.118 mmol) in DCE, add dimethylamine (3.87M in THF, 40 μL)followed by 1 drop of HOAc. Stir the mixture overnight, and then dilutewith DCM and wash (2×) with 10% NaOH solution. Concentrate under reducedpressure and add EtOAc, followed by a solution of HCl in ether.Concentrate and triturate the hydrochloride salt with fresh EtOAc toafford the title compound. ¹H NMR (400 MHz, CDCl₃): δ1.31 (d, 3H, J=6.23Hz, CH₃), 1.39 (d, 3H, J=6.6 Hz, CH3), 1.68 (m, 1H), 1.91 (m, 1H), 2.05(m, 5H), 2.68 (s, 6H, N(CH₃)₂)_(,) 2.9-3.1 (m, 4H), 3.65 (m, 1H),3.63-3.9 (m, 4H), 4.34 (m, 2H), 4.67 (br s, 1H), 6.20 (s, 1H), 7.18 (t,1H), 7.55 (s, 1H), 7.88 (m, 1H), 8.06 (s, 2H).

4.2-(5-chloro-6-((R)-4-(6-(3-chloro-4-fluorophenyl)-5-fluoro-2-((R)-2-methylpyrrolidin-1-yl)pyrimidin-4-yl)-3-methylpiperazin-1-yl)pyridin-3-yl)-N,N-dimethylethanamine

To a solution of2-(5-chloro-6-((R)-4-(6-(3-chloro-4-fluorophenyl)-2-((R)-2-methylpyrrolidin-1-yl)pyrimidin-4-yl)-3-methylpiperazin-)1-yl)pyridin-3-yl)-N,N-dimethylethanamine(50 mg, 0.087 mmol) in acetonitrile, add SELECTFLUOR® (93 mg, 0.26mmol). Heat the mixture at 150° C. for 10 min in microwave reactor, coolto room temperature, concentrate, and partition between EtOAc andsaturated NaHCO₃ solution. Concentrate under reduced pressure and purifywith flash chromatography to afford the title compound.

O.6-{4-[6-(4-Fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinicacid 1. 1-(5-Bromo-3-methyl-pyridin-2-yl)-3-(R)-methyl-piperazine

Heat a solution of 2,5-dibromo-3-methyl-pyridine (Chontech Inc.,Waterford, Conn.) (2.0 g, 7.97 mmol), (R)-2-methyl-piperazine(ChemPacific Corp., Baltimore, Md.; 3.2 g, 31.9 mmol) in DMA at 130° C.for 16 h. Partition the reaction mixture between water and EtOAc. Washthe EtOAc layer with water (1×) and brine (1×), dry (Na₂SO₄) andconcentrate under reduced pressure to give1-(5-bromo-3-methyl-pyridin-2-yl)-3-(R)-methyl-piperazine as a solid.

2. 2,4-dichloro-6-(4-fluorophenyl)pyrimidine

Dissolve 4-fluorobromobenzene (8.75 g, 0.05 moles) in anhydrous ether(80 mL) under nitrogen atmosphere and cool to −78° C. Add dropwise 1.6 Mn-BuLi (34 mL, 0.055 moles) and stir at −78° C. for 45 min. Dissolve2,4-dichloropyrimidine (7.45 g, 0.05 moles) in Et₂O (100 mL) and adddropwise to the reaction mixture. Warm the reaction mixture to −30° C.and stir at this temperature for 30 min followed by 0° C. for 30 min.Quench the reaction mixture with AcOH (3.15 mL, 0.055 moles) and water(0.5 mL, 0.027 moles) dissolved in THF (5.0 mL). Add dropwise a THF (40mL) solution of DDQ (11.9 g, 0.053 moles) to the reaction mixture. Bringthe reaction mixture to room temperature and stir at room temperaturefor 30 min. Cool the reaction mixture to 0° C., add 3.0 N aq. NaOH (35mL) and stir for 30 min. Decant the organic layer from the reactionmixture and wash the brown solid with Et₂O (3×100 mL). Combine theorganic layers, wash several times with saturated NaCl solution and drywith MgSO₄. Filter and evaporate under vacuum to afford a brown coloredsolid. Purify by flash column chromatography using 5% EtOAc/hexane toafford the title product as a white solid.

3.4-[4-(5-Bromo-3-methyl-pyridin-2-yl)-2-(R)-methyl-piperazin-1-yl]-2-chloro-6-(4-fluoro-phenyl)-pyrimidine

Heat a mixture of 2,4-dichloro-6-(4-fluoro-phenyl)-pyrimidine (6.0 g,24.7 mmol), 1-(5-bromo-3-methyl-pyridin-2-yl)-3-(R)-methyl-piperazine(7.0 g, 25.9 mmol) and K₂CO₃ (6.8 g, 49.4 mmol) in DMA at 60° C. for 16h. Partition the mixture between EtOAc and water, dry (Na₂SO₄) theorganic layer and concentrate under reduced pressure. Purify with flashsilica gel column eluting with 15% EtOAc/hexanes. Concentrate underreduced pressure to give the title compound.

4.4-[4-(5-Bromo-3-methyl-pyridin-2-yl)-2-(R)-methyl-piperazin-1-yl]-6-(4-fluoro-phenyl)-2-(2-(R)-methyl-pyrrolidin-1-yl)-pyrimidine

Heat a mixture of4-[4-(5-bromo-3-methyl-pyridin-2-yl)-2-(R)-methyl-piperazin-1-yl]-2-chloro-6-(4-fluoro-phenyl)-pyrimidine(7.7 g, 16.2 mmol), (R)-2-methylpyrrolidine hydrobromide [preparedessentially as described by Nijhuis et. al. (1989) J. Org. Chem.54(1):209] (3.5 g, 21.1 mmol) and K₂CO₃ (5.1 g, 37.3 mmol) in DMA at110° C. for 16 h. Partition the mixture between EtOAc and water, dry(Na₂SO₄) the organic layer and concentrate under reduced pressure.Purify with flash silica gel column eluting with 10% EtOAc/hexanes.Concentrate under reduced pressure to give the title compound.

5.6-{4-[6-(4-Fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinonitrile

To a mixture of4-[4-(5-bromo-3-methyl-pyridin-2-yl)-2-(R)-methyl-piperazin-1-yl]-6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidine(700 mg, 1.33 mmol) and Zn(CN)₂ (94 mg, 0.799 mmol) in DMF, addPd(PPh₃)₄ (77 mg, 0.067 mmol). Purge the reaction mixture for 10 minwith dry N₂. Heat the stirring reaction mixture overnight at 80° C.,cool to room temperature and partition between water and EtOAc. Dry thesolution (Na₂SO₄), concentrate under reduced pressure. Purify theresidue by flash column eluting with EtOAc-Hexanes (1:1) to afford thetitle compound as a white solid.

6.6-{4-[6-(4-Fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinicacid

Heat a solution of6-{4-[6-(4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-methyl-nicotinonitrile(100 mg, 0.212 mmol) in 12 M HCl for 3 hours at 90° C. Concentrate themixture under reduced pressure. Add a small amount of water, adjust thepH to 6-7, and collect the resulting white precipitate to afford thetitle compound as a off-white solid. ¹H NMR (300 MHz, DMSO-d₆): δ 1.24(m, 6H, 2×CH₃))_(;) 1.61 (m, 1H,); 1.84 (m, 1H); 1.98 (m, 2H); 2.34 (s,3H, Ar—CH₃); 2.91 (m, 1H); 3.08 (m, 1H); 3.26 (m, 2H); 3.56 (m, 2H);3.74 (m, 1H); 4.21 (m, 1H); 4.35 (m, 1H); 4.74 (m, 1H); 6.57 (s, 1H);7.26 (m, 2H); 7.91 (d, 1H, J=3 Hz); 8.15 (m, 2H); 8.60 (d, 1H, J=3 Hz).

Example 3 Additional Representative Substituted BiarylPiperazinyl-Pyridine Analogues

Using routine modifications, the starting materials may be varied andadditional steps employed to produce other compounds provided herein.Compounds listed in Tables I and II are prepared using such methods. A“*” in the column labeled “IC₅₀” in Table I indicates that the IC₅₀determined as described in Example 6 is 1 micromolar or less. Massspectroscopy data (column labeled “MS”) in Table I is obtained asdescribed above and is provided as M+1. Retention times are presented inmin.

TABLE I Representative Substituted Biaryl Piperazinyl-Pyridine AnaloguesRet. MS Compound Name IC₅₀ Time (M + 1) 1

2-{4-[6-(3-Chloro-4- fluoro-phenyl)-2- morpholin-4-yl- pyrimidin-4-yl]-piperazin-1-yl}-nicotinic acid ethyl ester * 1.27 527.30 2

2-{4-[6-(3-Chloro-4- fluoro-phenyl)-2- morpholin-4-yl- pyrimidin-4-yl]-piperazin-l-yl}- nicotinamide * 1.14 498.29 3

2-{4-[6-(3-Chloro-4- fluoro-phenyl)-2- morpholin-4-yl- pyrimidin-4-yl] -piperazin-1-yl}- nicotinic acid * 1.17 499.28 4

4-{4-(3-Chloro-4-fluoro- phenyl)-6-[4-(3-nitro-pyridin-2-yl)-piperazin-1- yl]-pyrimidin-2-yl}- morpholine * 1.26 500.275

4-(3-Chloro-4-fluoro- phenyl)-6-[4-(3-methyl- 5-nitro-pyridin-2-yl)-piperazin-1-yl]-2-(2- methyl-pyrrolidin-1-yl)- pyrimidine * 1.26 512.366

[2-(6-{4-[6-(3-Chloro-4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-5-methyl- pyridin-3-yloxy)-ethyl]-dimethyl-amine * 1.14 554.47 7

3-Chloro-2-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}- isonicotinic acid * 1.22 531.33 8

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3- (R)-methyl-piperazin-1- yl}-nicotinic acid methylester * 1.31 559.19 9

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3- (R)-methyl-piperazin-l- yl}-nicotinic acid * 1.27545.17 10

(5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-methanol * 1.23517.18 11

(5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-ylmethyl)-methyl-amine * 1.16 530.23 12

C-(5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)-2-(2-methyl-pyrrolidin-l- yl)-pyrimidin-4-yl]- piperazin-1-yl}-pyridin-3-yl)-methylamine * 1.15 516.20 13

(5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3- (R)-methyl-piperazin-1- yl}-pyridin-3-yl)-methanol * 1.25 531.21 14

4-(3-Chloro-4-fluoro- phenyl)-6-[4-(3-methyl- 5-nitro-pyridin-2-yl)-piperazin-l-yl]-2-(4- propyl-piperazin-1-yl)- pyrimidine * 1.26 555.2615

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-l-yl}- nicotinamide * 1.22 530.19 16

(5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(4-propyl-piperazin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-methanol * 1.23560.24 17

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(4-propyl-piperazin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid * 1.26 574.19 18

5-Chloro-6-{4-[6-(3- chloro-4- fluoro-phenyl)- 2-(2-methyl-pyrrolidin-l-yl)-pyrimidin-4-yl]-3- (R)-methyl-piperazin-1- yl}-nicotinic acid ethylester * 1.33 573.22 19

(5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-methyl-pyrrolidin-l-yl)-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-ylmethyl)-dimethyl-amine * 1.15 544.24 20

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid methyl ester * 1.29545.20 21

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid * 1.26 531.21 22

4-(3-Chloro-4-fluoro- phenyl)-6-[4-(6-methoxy- 3-nitro-pyridin-2-yl)-piperazin-1-yl]-2-(2- methyl-pyrrolidin-1-yl)- pyrimidine * 1.26 528.2123

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(4-propyl-piperazin-l-yl)-pyrimidin-4-yl] - piperazin-1-yl}-nicotinic acid ethyl ester * 1.32602.26 24

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3- (R)-methyl-piperazin-l- yl}-nicotinamide * 1.24544.19 25

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid * 1.23 497.22 26

(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-methanol * 1.2 483.23 27

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- nicotinic acid ethylester * 1.28 539.30 28

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2- pyrrolidin-1-yl-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- nicotinic acid ethylester * 1.28 525.27 29

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- nicotinic acid * 1.23511.27 30

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2- pyrrolidin-1-yl-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- nicotinic acid * 1.22497.25 31

(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- pyridin-3-yl)-methanol *1.21 497.28 32

(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2- pyrrolidin-1-yl-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- pyridin-3-yl)-methanol *1.19 483.26 33

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- nicotinamide * 1.2 510.2834

6-{4-[6-(4-Fluoro- phenyl)-2-(2-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinic acid *1.2 491.31 35

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2- pyrrolidin-1-yl-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- nicotinamide 36

6-{4-[6-(4-Fluoro- phenyl)-2-(2-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-5-methyl- nicotinic acid * 1.18 477.3037

(6-{4-[6-(4-Fluoro- phenyl)-2-(2-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-pyridin-3-yl)-methanol * 1.13 477.34 38

(6-{4-[6-(4-Fluoro- phenyl)-2-(2-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-5-methyl- pyridin-3-yl)-methanol * 1.11463.32 39

(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-acetic acid * 1.21511.26 40

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}- nicotinamide * 1.18 496.26 41

5-Chloro-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-phenyl-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid ethyl ester *1.31 521.31 42

5-Chloro-6-{4-[6-(5- chloro-pyridin-3-yl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- nicotinic acid ethylester * 1.34 556.28 43

5-Chloro-6-{4-[6-(2- isopropyl-pyridin-4-yl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3- (R)-methyl-piperazin-1- yl}-nicotinic acid ethylester * 1.34 564.36 44

6-{4-[6-(4-Fluoro- phenyl)-2-(2-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinamide *1.17 490.34 45

6-{4-[6-(4-Fluoro- phenyl)-2-(2-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-5-methyl- nicotinamide * 1.15 476.32 46

5-Chloro-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-m-tolyl-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid ethyl ester * 1.3535.30 47

(5-Chloro-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-phenyl-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-methanol * 1.23479.26 48

(5-Chloro-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-m-tolyl-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-methanol * 1.25493.27 49

(5-Chloro-6-{4-[6-(5- chloro-pyridin-3-yl)-2-(2-methyl-pyrrolidin-1-yl)- pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-pyridin-3-yl)-methanol * 1.24 514.22 50

(5-Chloro-6-{4-[6-(2- isopropyl-pyridin-4-yl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3- (R)-methyl-piperazin-1- yl}-pyridin-3-yl)-methanol * 1.24 522.31 51

5-Chloro-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-m-tolyl-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid * 1.27 507.25 52

5-Chloro-6-{4-[6-(5- chloro-pyridin-3-yl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- nicotinic acid * 1.28528.20 53

5-Chloro-6-{4-[6-(2- isopropyl-pyridin-4-yl)- 2-(2-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3- (R)-methyl-piperazin-1- yl}-nicotinic acid * 1.28536.28 54

5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6-phenyl-pyrimidin-4-yl]-piperazin-1-yl}- nicotinic acid ethyl ester * 1.26 487.30 55

5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6-m-tolyl-pyrimidin-4-yl]-piperazin-1-yl}- nicotinic acid ethyl ester * 1.28 501.31 56

5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6-pyridin-4-yl-pyrimidin-4-yl]-piperazin-1-yl}- nicotinic acid ethyl ester * 1.24 488.31 57

6-{4-[6-(2-Isopropyl- pyridin-4-yl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-5-methyl - nicotinic acid ethyl ester *1.29 530.26 58

5-Methyl-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-phenyl-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid ethyl ester *1.27 501.31 59

5-Methyl-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-m-tolyl-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid ethyl ester * 1.29515.32 60

5-Methyl-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-pyridin-4-yl-pyrimidin-4- yl] -piperazin-1-yl}- nicotinic acid ethylester * 1.26 502.32 61

6-{4-[6-(2-Isopropyl- pyridin-4-yl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinic acidethyl ester * 1.3 544.36 62

6-{4-[2-Dimethylamino- 6-(4-fluoro-phenyl)- pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5- methyl-nicotinic acid ethyl ester * 1.24479.30 63

(5-Methyl-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-phenyl-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-methanol * 1.13459.31 64

(5-Methyl-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-m-tolyl-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-methanol * 1.17473.33 65

(5-Methyl-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-pyridin-4-yl-pyrimidin-4- yl]-piperazin-1-yl}- pyridin-3-yl)-methanol *1.1 460.32 66

(6-{4-[6-(2-Isopropyl- pyridin-4-yl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-pyridin-3-yl)-methanol * 1.16 502.34 67

(6-{4-[2-Dimethylamino- 6-(4-fluoro-phenyl)- pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5- methyl-pyridin-3-yl)- methanol * 1.09 437.2868

5-Methyl-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-phenyl-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid * 1.22 473.30 69

5-Methyl-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-m-tolyl-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid * 1.24 487.32 70

5-Methyl-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-pyridin-4-yl-pyrimidin-4- yl]-piperazin-1-yl}- nicotinic acid * 1.2474.31 71

(5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6-phenyl-pyrimidin-4-yl]-piperazin-1-yl}- pyridin-3-yl)-methanol * 1.07 445.31 72

(5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6-m-tolyl-pyrimidin-4-yl]-piperazin-1-yl}- pyridin-3-yl)-methanol * 1.10 459.33 73

(5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6-pyridin-4-yl-pyrimidin-4-yl]-piperazin-1-yl}- pyridin-3-yl)-methanol * 1.03 446.32 74

(6-{4-[6-(2-Isopropyl- pyridin-4-yl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-5-methyl- pyridin-3-yl)-methanol * 1.09488.39 75

5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6-phenyl-pyrimidin-4-yl]-piperazin-1-yl}- nicotinic acid * 1.2 459.29 76

5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6-m-tolyl-pyrimidin-4-yl]-piperazin-1-yl}- nicotinic acid * 1.22 473.29 77

5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6-pyridin-4-yl-pyrimidin-4-yl]-piperazin-1-yl}- nicotinic acid * 1.17 460.29 78

6-{4-[6-(2-Isopropyl- pyridin-4-yl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-5-methyl- nicotinic acid * 1.22 502.3279

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}- nicotinic acid ethyl ester * 1.34 490.17 80

6-{4-[6-(2-Isopropyl- pyridin-4-yl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinic acid *1.24 516.35 81

6-{4-[2-Dimethylamino- 6-(4-fluoro-phenyl) - pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5- methyl-nicotinic acid * 1.18 451.27 82

5-Methyl-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-phenyl-pyrimidin-4-yl]- piperazin-1-yl}- nicotinamide * 1.18 472.30 83

6-{4-[2-Dimethylamino- 6-(4-fluoro-phenyl)- pyrimidin-4-yl]-3-(R)-mcthyl-piperazin-1-yl}-5- methyl-nicotinamide * 1.14 450.29 84

6-[4-(2-Dimethylamino- 6-phenyl-pyrimidin-4- yl)-3-(R)-methyl-piperazin-1-yl]-5-methyl- nicotinamide * 1.13 432.25 85

6-[4-(2-Dimethylamino- 6-pyridin-4-yl-pyrimidin- 4-yl)-3-(R)-methyl-piperazin-1-yl]-5-methyl- nicotinamide * 1.1 433.26 86

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}- nicotinamide * 1.22 461.11 87

5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6- phenyl-pyrimidin-4-yl]-piperazin-1-yl}- nicotinamide * 1.17 458.28 88

5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6-m-tolyl-pyrimidin-4-yl]-piperazin-1-yl}- nicotinamide * 1.19 472.30 89

5-Methyl-6-{4-[2-(2- methyl-pyrrolidin-1-yl)- 6-pyridin-4-yl-pyrimidin-4-yl]-piperazin-1-yl}- nicotinamide * 1.14 459.30 90

6-{4-[6-(2-Isopropyl- pyridin-4-yl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-5-methyl- nicotinamide * 1.19 501.33 91

6-{4-[6-(4-fluorophenyl)- 2-(2-methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-5- methylpyridine-3- carboximidamide * 1.12 238.15 92

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2-(R)- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid * 1.24 497.26 93

(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2-(R)- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-acetic acid * 1.23511.27 94

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2-(R)- methyl-pyrrolidin-1-yl) -pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl} - nicotinic acid * 1.25511.24 95

6-{4-[6-(4-Fluoro- phenyl)-2-(2-(R)-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinic acid *1.21 491.29 96

5-Methyl-6-{3-(R)- methyl-4-[2-(2-(R)- methyl-pyrrolidin-1-yl)-6-phenyl-pyrimidin-4- yl]-piperazin-1-yl}- nicotinic acid * 1.21 473.2997

2-{4-[6-(3-Chloro-4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}- pyrimidine-5-carboxylic acid amide *1.22 497.26 98

(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2-(R)- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- pyridin-3-yl)-methanol *1.25 497.28 99

6-{4-[6-(4-Fluoro- phenyl)-2-(2-(S)-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinic acid *1.24 491.31 100

(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2-(R)- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-pyridin-3-yl)-acetonitrile * 1.25 506.28 101

(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2-(R)- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- pyridin-3-yl)-aceticacid * 1.26 525.29 102

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-(R)-methyl-pyrrolidin-l-yl)- pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-nicotinamide * 1.27 544.29 103

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-(R)-methyl-pyrrolidin-l-yl)- pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-nicotinic acid * 1.27 545.22 104

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(3-(R)- methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- nicotinic acid * 1.21527.29 105

(6-{4-[6-(4-Fluoro- phenyl)-2-(2-(R)-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-pyridin-3-yl)-acetic acid * 1.12 505.35 106

N-Cyclopropyl-6-{4-[6- (4-fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-5-methyl- nicotinamide * 1.17 516.38107

N-cyclobutyl-6-{4-[6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1-yl)pyrimidin-4- yl]piperazin-1-yl}-5- methylnicotinamide * 1.2 530.39108

N-Cyclopentyl-6-{4-[6- (4-fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-5-methyl- nicotinamide * 1.22 544.41109

6-{4-[6-(4-fluorophenyl)- 2-(2-methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-N-(2- methoxyethyl)-5- methylnicotinamide * 1.17534.39 110

6-{4-[6-(4-fluorophenyl)- 2-(2-methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-N-(3- methoxypropyl)-5- methylnicotinamide * 1.18548.40 111

6-{4-[6-(4-fluorophenyl)- 2-(2-methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-5- methyl-N- (tetrahydrofuran-2-ylmethyl)nicotinamide * 1.18 560.41 112

N-(3-ethoxypropyl)-6-{4- [6-(4-fluorophenyl)-2-(2- methylpyrrolidin-1-yl)pyrimidin-4- yl]piperazin-1-yl}-5- methylnicotinamide * 1.2 562.43113

N-benzyl-6-{4-[6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1-yl)pyrimidin-4- yl]piperazin-1-yl}-5- methylnicotinamide * 1.22 566.40114

N-(4-fluorobenzyl)-6-{4- [6-(4-fluorophenyl)-2-(2- methylpyrrolidin-1-yl)pyrimidin-4- yl]piperazin-1-yl}-5- methylnicotinamide * 1.22 584.40115

6-{4-[6-(4-fluorophenyl)- 2-(2-methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-N-(4- methoxybenzyl)-5- methylnicotinamide * 1.22596.42 116

N-[3- (dimethylamino)propyl]- 6-{4-[6-(4-fluorophenyl)-2-(2-methylpyrrolidin-1- yl)pyrimidin-4- yl]piperazin-1-yl}-5-methylnicotinamide * 1.11 561.46 117

6-{4-[6-(4-fluorophenyl)- 2-(2-methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-N-(2- methoxy-1-methylethyl)- 5-methylnicotinamide *1.18 548.41 118

6-{4-[6-(4-fluorophenyl)- 2-(2-methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-5- methyl-N-(2- thienylmethyl) nicotinamide * 1.21572.37 119

N-(2-ethoxyethyl)-6-{4- [6-(4-fluorophenyl)-2-(2- methylpyrrolidin-1-yl)pyrimidin-4- yl]piperazin-1-yl}-5- methylnicotinamide * 1.18 548.41120 6-{4-[6-(4-fluorophenyl)- 2-(2-methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-5- methyl-N-(2,2,2- trifluoroethyl)nicotinamide * 1.2558.36 121

N-benzyl-6-{4-[6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1-yl)pyrimidin-4- yl]piperazin-1-yl}-N,5- dimethylnicotinamide * 1.22580.43 122

4-(4-fluorophenyl)-2-(2- methylpyrrolidin-1-yl)-6- {4-[3-methyl-5-(pyrrolidin-1- ylcarbonyl)pyridin-2- yl]piperazin-1- yl}pyrimidine *1.18 530.40 123

4-(4-fluorophenyl)-6-{4- [3-methyl-5-(piperidin-1- ylcarbonyl)pyridin-2-yl]piperazin-1-yl}-2-(2- methylpyrrolidin-1- yl)pyrimidine * 1.2 544.42124

4-[(6-{4-[6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-5- methylpyridin-3- yl)carbonyl]morpholine * 1.16546.40 125

4-(4-fluorophenyl)-6-(4- {3-methyl-5-[(4- methylpiperidin-1-yl)carbonyl]pyridin-2- yl}piperazin-1-yl)-2-(2- methylpyrrolidin-1-yl)pyrimidine * 1.23 558.44 126

l-[(6-{4-[6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-5- methylpyridin-3- yl)carbonyl]azepane * 1.22 558.44127

4-[(6-{4-[6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-5- methylpyridin-3- yl)carbonyl] thiomorpholine *1.19 562.38 128

4-(4-fluorophenyl)-6-(4- {3-methyl-5-[(2- methylpiperidin-1-yl)carbonyl]pyridin-2- yl}piperazin-1-yl)-2-(2- methylpyrrolidin-1-yl)pyrimidine * 1.22 558.44 129

N-cyclohexyl-6-{4-[6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1-yl)pyrimidin-4- yl]piperazin-1-yl}-N,5- dimethylnicotinamide * 1.24572.45 130

4-(4-fluorophenyl)-6-(4- {3-methyl-5-[(4- phenylpiperidin-1-yl)carbonyl]pyridin-2- yl}piperazin-1-yl)-2-(2- methylpyrrolidin-1-yl)pyrimidine * 1.26 620.48 131

4-(4-fluorophenyl)-6-(4- {3-methyl-5-[(4- methylpiperazin-1-yl)carbonyl]pyridin-2- yl}piperazin-1-yl)-2-(2- methylpyrrolidin-1-yl)pyrimidine * 1.1 559.44 132

4-(4-fluorophenyl)-6-[4- (5-{[4-(2- methoxyethyl)piperazin-1-yl]carbonyl}-3- methylpyridin-2- yl)piperazin-1-yl]-2-(2-methylpyrrolidin-1- yl)pyrimidine * 1.11 603.48 133

4-(4-{5-[(4- cyclopentylpiperazin-1- yl)carbonyl]-3- methylpyridin-2-yl}piperazin-1-yl)-6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1-yl)pyrimidine * 1.12 613.51 134

4-(4-{5-[(4- acetylpiperazin-1- yl)carbonyl]-3- methylpyridin-2-yl}piperazin-1-yl)-6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1-yl)pyrimidine * 1.14 587.44 135

4-(4-fluorophenyl)-6-(4- {3-methyl-5-[(2- methylpyrrolidin-1-yl)carbonyl]pyridin-2- yl}piperazin-1-yl)-2-(2- methylpyrrolidin-1-yl)pyrimidine * 1.2 544.42 136

N-[2- (dimethylamino)ethyl]-6- {4-[6-(4-fluorophenyl)-2-(2-methylpyrrolidin-1- yl)pyrimidin-4- yl]piperazin-1-yl}-N,5-dimethylnicotinamide * 1.1 561.46 137

N-[3- (dimethylamino)propyl]- 6-{4-[6-(4-fluorophenyl)-2-(2-methylpyrrolidin-1- yl)pyrimidin-4- yl]piperazin-1-yl}-N,5-dimethylnicotinamide * 1.11 575.48 138

6-{4-[6-(4-fluorophenyl)- 2-(2-methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-N,5- dimethyl-N-(1- methylpyrrolidin-3-yl)nicotinamide * 1.11 573.47 139

l-[(6-{4-[6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1- yl)pyrimidin-4-yl]piperazin-1-yl}-5- methylpyridin-3- yl)carbonyl]-N,N-dimethylpyrrolidin-3 amine * 1.1 573.47 140

N-{l-[(6-{4-[6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1-yl)pyrimidin-4- yl]piperazin-1-yl}-5- methylpyridin-3-yl)carbonyl]pyrrolidin-3- yl}acetamide * 1.15 587.45 141

4-(4-fluorophenyl)-6-(4- {5-[(4-methoxypiperidin- 1-yl)carbonyl]-3-methylpyridin-2- yl}piperazin-1-yl)-2-(2- methylpyrrolidin-1-yl)pyrimidine * 1.18 574.45 142

4-(4-{5-[(4,4- difluoropiperidin-1- yl)carbonyl]-3- methylpyridin-2-yl}piperazin-1-yl)-6-(4- fluorophenyl)-2-(2- methylpyrrolidin-1-yl)pyrimidine * 1.2 580.41 143

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(3-(R)- methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- nicotinamide * 1.17526.39 144

6-{4-[6-(4-Fluoro- phenyl)-2-(2-(R)-methyl- pyrrolidin-l-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-pyridine-2-carboxylic acid * 1.18 491.40 145

6-{4-[6-(4-Fluoro- phenyl)-2-(2-(R)-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-pyridine-2-carboxylic acid amide * 1.23 490.22 146

(5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-(R)-methyl-pyrrolidin-1-yl)- pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-pyridin-3-yl)-methanol * 1.29 531.15 147

2-(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-propan-2-ol * 1.28511.26 148

l-(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-ethanol * 1.26 497.23149

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-(R)-methyl-pyrrolidin-1-yl)- pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-N-(2-hydroxy-ethyl)- nicotinamide * 1.28 588.24 150

6-{4-[6-(4-Fluoro- phenyl)-2-(3-(R)-methyl- morpholin-4-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinic acid *1.25 507.28 151

6-{4-[6-(4-Fluoro- phenyl)-2-(2-(S)- hydroxymethyl- pyrrolidin-l-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinic acid *1.23 507.27 152

2-(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-pyridin- 3-yl)-acetamide * 1.22 510.16153

6-{4-[6-(4-Fluoro- phenyl)-2-(isopropyl- methyl-amino)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinic acid *1.25 479.21 154

6-{4-[6-(3-Chloro-4- fluoro-phenyl)-2-(2-(R)- methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinic acid *1.29 525.17 155

(5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2-(S)- hydroxymethyl-pyrrolidin-1-yl)- pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-pyridin-3-yl)-(2-(S)- hydroxymethyl- pyrrolidin-1-yl)- methanone * 1.24610.24 156

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2-(S)- hydroxymethyl-pyrrolidin-1-yl)- pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-nicotinic acid * 1.27 527.16 157

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2- (isopropyl-methyl-amino)-pyrimidin-4-yl]- 3-(R)-methyl-piperazin- 1-yl}-nicotinic acid *1.28 499.16 158

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(2-(S)- hydroxymethyl-pyrrolidin-1-yl)- pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid * 1.25513.19 159

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2-(3-(R)- methyl-morpholin-4-yl)-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid * 1.28 513.17 160

5-Chloro-6-{4-[6-(4- fluoro-phenyl)-2- (isopropyl-methyl-amino)-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid * 1.26 485.19 161

6-{4-[6-(3-Chloro-4- fluoro-phenyl)-2- (isopropyl-methyl-amino)-pyrimidin-4-yl] - piperazin-1-yl}-5-methyl- nicotinic acid * 1.28499.21 162

4-(4-Fluoro-phenyl)-6-[2- (R)-methyl-4-(3-methyl-5-oxazol-2-yl-pyridin-2- yl)-piperazin-1-yl]-2-(2-(R)-methyl-pyrrolidin-1- yl)-pyrimidine * 1.28 514.27 163

5-Chloro-6-{4-[2-(2-(S)- hydroxymethyl- pyrrolidin-1-yl)-6-phenyl-pyrimidin-4-yl]- 3-(R)-methyl-piperazin- 1-yl}-nicotinic acid *1.26 509.21 164

4-(4-Fluoro-phenyl)-6- {2-(R)-methyl-4-[3- methyl-5-(1H-tetrazol-5-yl)-pyridin-2-yl]- piperazin-1-yl}-2-(2-(R)- methyl-pyrrolidin-1-yl)-pyrimidine * 1.24 515.27 165

5-Chloro-6-{4-[2-(2-(S)- hydroxymethyl- pyrrolidin-1-yl)-6-phenyl-pyrimidin-4-yl]- 3-(R)-methyl-piperazin- 1-yl}-nicotinamide *1.23 508.22 166

5-Chloro-N-(2-hydroxy- ethyl)-6-{4-[2-(2-(S)- hydroxymethyl-pyrrolidin-1-yl)-6- phenyl-pyrimidin-4-yl]- 3-(R)-methyl-piperazin-1-yl}-nicotinamide * 1.23 552.26 167

5-Chloro-N-(2-hydroxy- ethyl)-6-{3-(R)-methyl-4- [2-(2-(R)-methyl-pyrrolidin-1-yl)-6- phenyl-pyrimidin-4-yl]- piperazin-1-yl}-nicotinamide * 1.25 536.29 168

5-Methyl-6-{3-(R)- methyl-4-[2-(2-methyl- pyrrolidin-1-yl)-6-p-tolyl-pyrimidin-4-yl]- piperazin-1-yl}-nicotinic acid * 1.22 488.30 169

4-(4-Fluoro-phenyl)-6- {4-[5-(1H-imidazol-2- yl)-3-methyl-pyridin-2-yl]-2-(R)-methyl- piperazin-1-yl}-2-(2-(R)- methyl-pyrrolidin-1-yl)-pyrimidine * 1.18 257.14 170

6-{4-[6-(4-Fluoro- phenyl)-2-(2-(R)-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(S)- methyl-piperazin-1-yl}-5- methyl-nicotinic acid *1.17 491.36 171

N-(2-Hydroxy-ethyl)-5- methyl-6-{3-(R)-methyl- 4-[6-(6-methyl-pyridin-3-yl)-2-(2-methyl- pyrrolidin-1-yl)- pyrimidin-4-yl]- piperazin-1-yl}-nicotinamide * 1.1 531.42 172

6-{4-[2-(4-Fluoro- phenyl)-6-(2-(R)-methyl- pyrrolidin-1-yl)-pyridin-4-yl]-3-(R)-methyl- piperazin-1-yl}-5-methyl- nicotinic acid * 1.16490.39 173

(5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-piperidin-1-yl-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}- pyridin-3-yl)-methanol *1.24 531.23 174

5-Chloro-6-{4-[6-(3- chloro-4-fluoro-phenyl)- 2-(2-(R)-methyl-pyrrolidin-1-yl)- pyrimidin-4-yl]-2-(R)- methyl-piperazin-1-yl}-nicotinic acid ethyl ester * 1.29 573.22 175

6-{4-[2-(3-Chloro- phenyl)-6-(2-(R)-methyl- pyrrolidin-1-yl)-pyridin-4-yl]-3-(R)-methyl- piperazin-1-yl}-5-methyl- nicotinic acid * 1.2506.25 176

6-{4-[2-(4-Fluoro-3- methyl-phenyl)-6-(2-(R)- methyl-pyrrolidin-1-yl)-pyridin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinic acid *1.2 504.32 177

6-{4-[6-(4-Fluoro- phenyl)-2-(2-(R)-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]- piperazin-1-yl}-5- trifluoromethyl-nicotinic acid *1.21 531.21 178

6-{4-[2-(3-Chloro-4- fluoro-phenyl)-6-(2-(R)- methyl-pyrrolidin-1-yl)-pyridin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5- methyl-nicotinic acid *1.19 524.24 179

6-{4-[6-(4-Fluoro- phenyl)-2-(2-(R)-methyl- pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)- methyl-piperazin-1-yl}-5-trifluoromethyl-nicotinic acid * 1.22 545.24 180

6-{4-[5′-Chloro-6-(2-(R)- methyl-pyrrolidin-1-yl)-[2,3′]bipyridinyl-4-yl]-3- (R)-methyl-piperazin-1-yl}-5-methyl-nicotinic acid * 1.14 507.20 181

2-[6-((3R)-4-{6-(4- fluorophenyl)-2-[(2R)-2- methyl-1-pyrrolidinyl]-4-pyrimidinyl}-3-methyl-1- piperazinyl)-5-methyl-3- pyridinyl]ethanol *1.19 491.24 182

2-{[6-((3R)-4-{6-(3- chloro-4-fluorophenyl)-2- [(2R)-2-methyl-1-pyrrolidinyl]-4- pyrimidinyl}-3-methyl-1- piperazinyl)-5-methyl-3-pyridinyl]oxy}ethanol * 1.25 541.21 183

2-{[5-chloro-6-((3R)-4- {6-(3-chloro-4- fluorophenyl)-2-[(2R)-2-methylpyrrolidin-1- yl]pyrimidin-4-yl}-3- methylpiperazin-1-yl)pyridin-3- yl]oxy}ethanol * 1.29 561.16 184

2-{[6-((3R)-4-{6-(4- fluorophenyl)-2-[(2S)-2- (hydroxymethyl)pyrrolidin-1-yl]pyrimidin-4-yl}-3- methylpiperazin-1-yl)-5- methylpyridin-3-yl]oxy}ethanol * 1.2 523.40 185

2-[5-chloro-6-((3R)-4-{6- (3-chloro-4- fluorophenyl)-2-[(2R)-2-methylpyrrolidin-1- yl]pyrimidin-4-yl}-3- methylpiperazin-1-yl)pyridin-3-yl]ethanol * 1.2 545.23 186

2-[5-chloro-6-((3R)-4-{6- (3-chloro-4- fluorophenyl)-2-[(2R)-2-methylpyrrolidin-1- yl]pyrimidin-4-yl}-3- methylpiperazin-1-yl)pyridin-3-yl]-N,N- dimethylethanamine * 1.13 572.27 187

2-[5-chloro-6-((3R)-4-{6- (3-chloro-4- fluorophenyl)-2-[(2R)-2-methylpyrrolidin-1- yl]pyrimidin-4-yl}-3- methylpiperazin-1-yl)pyridin-3-yl]-N- methylethanamine * 1.23 558.48 188

6-((3R)-4-{6-(4- fluorophenyl)-2-[(2R)-2- methyl-1-pyrrolidinyl]-4-pyrimidinyl}-3-methyl-1- piperazinyl)-5- methylnicotinonitrile * 1.26471.33 189

5-fluoro-4-(4- fluorophenyl)-6-[(2R)-2- methyl-4-(3- methylpyridin-2-yl)piperazin-1-yl]-2- [(2R)-2- methylpyrrolidin-1- yl]pyrimidine * 1.26465.34 190

tert-butyl rel-{6-[(3R)-4- {6-(4-fluorophenyl)-2- [(2R)-2-methylpyrrolidin-1- yl]pyrimidin-4-yl}-3- methylpiperazin-1-yl]-5-methylpyridin-3- yl}acetate * 1.22 561.30 191

rel-(2E)-3-{5-chloro-6- [(3R)-4-{6-(3-chloro-4- fluorophenyl)-2-[(2R)-2-methylpyrrolidin-1- yl]pyrimidin-4-yl}-3- methylpiperazin-1-yl]pyridin-3-yl}prop-2- en-1-ol * 1.23 557.18 192

rel-2-({6-[(3R)-4-{6-(4- fluorophenyl)-2-[(2R)-2- methylpyrrolidin-1-yl]pyrimidin-4-yl}-3- methylpiperazin-1-yl]-5- methylpyridin-3-yl}oxy)ethanol * 1.15 507.26 193

rel-3-{5-chloro-6-[(3R)- 4-{6-(3-chloro-4- fluorophenyl)-2-[(2R)-2-methylpyrrolidin-1- yl]pyrimidin-4-yl}-3- methylpiperazin-1-yl]pyridin-3-yl}propan-1- ol * 1.32 559.41

TABLE II Additional Representative Substituted BiarylPiperazinyl-Pyridine Analogues Compound Name 194

5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-nicotinic acid 195

5-Chloro-6-{4-[6-(3-chloro-4-fluoro-phenyl)-2-(2-(S)-methyl-piperidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-nicotinic acid 196

5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(3-(S)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-nicotinic acid 197

5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-nicotinic acid 198

5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-nicotinic acid 199

5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-nicotinic acid 200

5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-nicotinamide 201

5-Chloro-6-{4-[6-(3-chloro-4-fluoro-phenyl)-2-(2-(S)-methyl-piperidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}- nicotinamide 202

5-Chloro-6-{4-[6-(4-cyano-phenyl)-2-(3-(S)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-nicotinamide 203

5-Chloro-6-{4-[6-(4-chloro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-nicotinamide 204

5-Chloro-6-{4-[6-(3,4-difluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-nicotinamide 205

5-Chloro-6-{4-[6-(4-fluoro-3-methyl-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}- nicotinamide 206

5-Chloro-6-{3-(S)-methyl-4-[6-(5-methyl-pyridin-3-yl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-piperazin-1-yl}- nicotinamide 207

5-Chloro-6-{4-[6-(2-isopropyl-pyridin-4-yl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}- nicotinamide 208

(6-{4-[6-(4-Fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-trifluoromethyl- pyridin-3-yl)-methanol 209

(6-{4-[6-(3-Chloro-4-fluoro-phenyl)-2-(2-(S)-methyl-piperidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-trifluoromethyl- pyridin-3-yl)-methanol 210

4-[6-[4-(5-Hydroxymethyl-3-trifluoromethyl-pyridin-2-yl)-2-(R)-methyl-piperazin-1-yl]-2-(3-(S)-methyl-morpholin-4-yl)-pyrimidin-4- yl]-benzonitrile 211

(6-{4-[6-(4-Chloro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-trifluoromethyl- pyridin-3-yl)-methanol 212

(6-{4-[6-(3,4-Difluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-5-trifIuoromethyl- pyridin-3-yl)-methanol 213

(6-{4-[6-(4-Fluoro-3-methyl-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-5- trifluoromethyl-pyridin-3-yl)-methanol214

(6-{4-[6-(5-Chloro-pyridin-3-yl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-5-trifluoromethyl- pyridin-3-yl)-methanol 215

(6-{4-[6-(2-Isopropyl-pyridin-4-yl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-trifluoromethyl- pyridin-3-yl)-methanol 216

(6-{4-[6-(4-Fluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-trifluoromethyl- pyridin-3-yl)-acetic acid 217

(6-{4-[6-(3-Chloro-4-fluoro-phenyl)-2-(2-(S)-methyl-piperidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-trifluoromethyl- pyridin-3-yl)-acetic acid 218

(6-{4-[6-(4-Cyano-phenyl)-2-(3-(S)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-5-trifluoromethyl- pyridin-3-yl)-acetic acid 219

(5-Chloro-6-{4-[6-(4-chloro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-pyridin-3-yl)- acetic acid 220

(6-{4-[6-(3,4-Difluoro-phenyl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-5-trifluoromethyl- pyridin-3-yl)-acetic acid221

(5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(3-(R)-methyl-morpholin-4-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-pyridin-3-yl)- acctic acid 222

(6-{4-[6-(5-Chloro-pyridin-3-yl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-5-trifluoromethyl- pyridin-3-yl)-acetic acid223

(5-Chloro-6-{4-[6-(2-isopropyl-pyridin-4-yl)-2-(2-(S)-methyl-pyrrolidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-pyridin-3- yl)-acetic acid 224

5-Chloro-6-{4-[6-(4-fluoro-phenyl)-2-(azetidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl- piperazin-1-yl}-nicotinicacid 225

5-Chloro-6-{4-[6-(4-chloro-phenyl)-2-(2-(S)-methyl-azetidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl}-nicotinic acid 226

5-Chloro-6-{4-[6-(3,4-difluoro-phenyl)-2-(2-(R)-methyl-azetidin-1-yl)-pyrimidin-4-yl]-3-(R)-methyl-piperazin-1-yl)-nicotinic acid 227

6-{4-[2-Azetidin-1-yl-6-(4-chloro-phenyl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-5-trifluoromethyl-nicotinamide 228

(6-{4-[6-(4-Fluoro-phenyl)-2-(2-(R)-methyl-azetidin-1-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-5-trifluoromethyl-pyridin-3- yl)-methanol 229

(6-{4-[6-(2-Isopropyl-pyridin-4-yl)-2-(2-(S)-methyl-azetidin-1-yl)-pyrimidin-4-yl]-3-(S)-methyl-piperazin-1-yl}-5-trifluoromethyl- pyridin-3-yl)-methanol

Example 4 VR1-Transfected Cells and Membrane Preparations

This Example illustrates the preparation of VR1-transfected cells andVR1-containing membrane preparations for use in capsaicin binding assays(Example 5).

A cDNA encoding full length human capsaicin receptor (SEQ ID NO:1, 2 or3 of U.S. Pat. No. 6,482,611) is subcloned in the plasmid pBK-CMV(Stratagene, La Jolla, Calif.) for recombinant expression in mammaliancells.

Human embryonic kidney (HEK293) cells are transfected with the pBK-CMVexpression construct encoding the full length human capsaicin receptorusing standard methods. The transfected cells are selected for two weeksin media containing G418 (400 μg/ml) to obtain a pool of stablytransfected cells. Independent clones are isolated from this pool bylimiting dilution to obtain clonal stable cell lines for use insubsequent experiments.

For radioligand binding experiments, cells were seeded in T175 cellculture flasks in media without antibiotics and grown to approximately90% confluency. The flasks were then washed with PBS and harvested inPBS containing 5 mM EDTA. The cells were pelleted by gentlecentrifugation and stored at −80° C. until assayed.

Previously frozen cells are disrupted with the aid of a tissuehomogenizer in ice-cold HEPES homogenization buffer (5 mM KCl5, 5.8 mMNaCl, 0.75 mM CaCl₂, 2 mM MgCl₂, 320 mM sucrose, and 10 mM HEPES pH7.4). Tissue homogenates are first centrifuged for 10 min at 1000×g (4°C.) to remove the nuclear fraction and debris, and then the supernatantfrom the first centrifugation is further centrifuged for 30 min at35,000×g (4° C.) to obtain a partially purified membrane fraction.Membranes are resuspended in the HEPES homogenization buffer prior tothe assay. An aliquot of this membrane homogenate is used to determineprotein concentration via the Bradford method (BIO-RAD Protein AssayKit, #500-0001, BIO-RAD, Hercules, Calif.).

Example 5 Capsaicin Receptor Binding Assay

This Example illustrates a representative assay of capsaicin receptorbinding that may be used to determine the binding affinity of compoundsfor the capsaicin (VR1) receptor.

Binding studies with [³H] Resiniferatoxin (RTX) are carried outessentially as described by Szallasi and Blumberg (1992) J. Pharmacol.Exp. Ter. 262:883-888. In this protocol, non-specific RTX binding isreduced by adding bovine alpha₁ acid glycoprotein (100 μg per tube)after the binding reaction has been terminated.

[³H] RTX (37 Ci/mmol) is synthesized by and obtained from the ChemicalSynthesis and Analysis Laboratory, National Cancer Institute-FrederickCancer Research and Development Center, Frederick, Md. [³H] RTX may alsobe obtained from commercial vendors (e.g., Amersham Pharmacia Biotech,Inc.; Piscataway, N.J.).

The membrane homogenate of Example 4 is centrifuged as before andresuspended to a protein concentration of 333 μg/ml in homogenizationbuffer. Binding assay mixtures are set up on ice and contain [³H]RTX(specific activity 2200 mCi/ml), 2 μl non-radioactive test compound,0.25 mg/ml bovine serum albumin (Cohn fraction V), and 5×10⁴-1×10⁵VR1-transfected cells. The final volume is adjusted to 500 μl (forcompetition binding assays) or 1,000 μl (for saturation binding assays)with the ice-cold HEPES homogenization buffer solution (pH 7.4)described above. Non-specific binding is defined as that occurring inthe presence of 1 μM non-radioactive RTX (Alexis Corp.; San Diego,Calif.). For saturation binding, [³H]RTX is added in the concentrationrange of 7-1,000 pM, using 1 to 2 dilutions. Typically 11 concentrationpoints are collected per saturation binding curve.

Competition binding assays are performed in the presence of 60 pM[³H]RTX and various concentrations of test compound. The bindingreactions are initiated by transferring the assay mixtures into a 37° C.water bath and are terminated following a 60 minute incubation period bycooling the tubes on ice. Membrane-bound RTX is separated from free, aswell as any alpha₁-acid glycoprotein-bound RTX, by filtration ontoWALLAC glass fiber filters (PERKIN-ELMER, Gaithersburg, Md.) which werepre-soaked with 1.0% PEI (polyethyleneimine) for 2 h prior to use.Filters are allowed to dry overnight then counted in a WALLAC 1205 BETAPLATE counter after addition of WALLAC BETA SCINT scintillation fluid.

Equilibrium binding parameters are determined by fitting the allostericHill equation to the measured values with the aid of the computerprogram FIT P (Biosoft, Ferguson, Mo.) as described by Szallasi, et al.(1993) J. Pharmacol. Exp. Ther. 266:678-683. Compounds provided hereingenerally exhibit K_(i) values for capsaicin receptor of less than 1 μM,100 nM, 50 nM, 25 nM, 10 nM, or 1 nM in this assay.

Example 6 Calcium Mobilization Assay

This Example illustrates representative calcium mobilization assays foruse in evaluating test compounds for agonist and antagonist activity.

Cells transfected with expression plasmids (as described in Example 4)and thereby expressing human capsaicin receptor are seeded and grown to70-90% confluency in FALCON black-walled, clear-bottomed 96-well plates(#3904, BECTON-DICKINSON, Franklin Lakes, N.J.). The culture medium isemptied from the 96 well plates and FLUO-3 AM calcium sensitive dye(Molecular Probes, Eugene, Oreg.) is added to each well (dye solution: 1mg FLUO-3 AM, 440 μL DMSO and 440 μl 20% pluronic acid in DMSO, diluted1:250 in Krebs-Ringer HEPES (KRH) buffer (25 mM HEPES, 5 mM KCl, 0.96 mMNaH₂PO₄, 1 mM MgSO₄, 2 mM CaCl₂, 5 mM glucose, 1 mM probenecid, pH 7.4),50 μl diluted solution per well). Plates are covered with aluminum foiland incubated at 37° C. for 1-2 h in an environment containing 5% CO₂.After the incubation, the dye is emptied from the plates, and the cellsare washed once with KRH buffer, and resuspended in KRH buffer.

Determination of Capsaicin EC₅₀

To measure the ability of a test compound to agonize or antagonize acalcium mobilization response in cells expressing capsaicin receptors tocapsaicin or other vanilloid agonist, the EC₅₀ of the agonist capsaicinis first determined. An additional 20 μl of KRH buffer and 1 μl DMSO isadded to each well of cells, prepared as described above. 100 μlcapsaicin in KRH buffer is automatically transferred by the FLIPRinstrument to each well. Capsaicin-induced calcium mobilization ismonitored using either FLUOROSKAN ASCENT (Labsystems; Franklin, Mass.)or FLIPR (fluorometric imaging plate reader system; Molecular Devices,Sunnyvale, Calif.) instruments. Data obtained between 30 and 60 secondsafter agonist application are used to generate an 8-point concentrationresponse curve, with final capsaicin concentrations of 1 nM to 3 μM.KALEIDAGRAPH software (Synergy Software, Reading, Pa.) is used to fitthe data to the equation:y=a*(1/(1+(b/x)^(c)))to determine the 50% excitatory concentration (EC₅₀) for the response.In this equation, y is the maximum fluorescence signal, x is theconcentration of the agonist or antagonist (in this case, capsaicin), ais the E_(max), b corresponds to the EC₅₀ value and c is the Hillcoefficient.Determination of Agonist Activity

Test compounds are dissolved in DMSO, diluted in KRH buffer, andimmediately added to cells prepared as described above. 100 nM capsaicin(an approximate EC₉₀ concentration) is also added to cells in the same96-well plate as a positive control. The final concentration of testcompounds in the assay wells is between 0.1 nM and 5 μM.

The ability of a test compound to act as an agonist of the capsaicinreceptor is determined by measuring the fluorescence response of cellsexpressing capsaicin receptors elicited by the compound as function ofcompound concentration. This data is fit as described above to obtainthe EC₅₀, which is generally less than 1 micromolar, preferably lessthan 100 nM, and more preferably less than 10 nM. The extent of efficacyof each test compound is also determined by calculating the responseelicited by a concentration of test compound (typically 1 μM) relativeto the response elicited by 100 nM capsaicin. This value, called Percentof Signal (POS), is calculated by the following equation:POS=100*test compound response/100nM capsaicin response

This analysis provides quantitative assessment of both the potency andefficacy of test compounds as human capsaicin receptor agonists.Agonists of the human capsaicin receptor generally elicit detectableresponses at concentrations less than 100 μM, or preferably atconcentrations less than 1 μM, or most preferably at concentrations lessthan 10 nM. Extent of efficacy at human capsaicin receptor is preferablygreater than 30 POS, more preferably greater than 80 POS at aconcentration of 1 μM. Certain agonists are essentially free ofantagonist activity as demonstrated by the absence of detectableantagonist activity in the assay described below at compoundconcentrations below 4 nM, more preferably at concentrations below 10 μMand most preferably at concentrations less than or equal to 100 μM.

Determination of Antagonist Activity

Test compounds are dissolved in DMSO, diluted in 20 μl KRH buffer sothat the final concentration of test compounds in the assay well isbetween 1 μM and 5 μM, and added to cells prepared as described above.The 96 well plates containing prepared cells and test compounds areincubated in the dark, at room temperature for 0.5 to 6 h. It isimportant that the incubation not continue beyond 6 h. Just prior todetermining the fluorescence response, 100 μl capsaicin in KRH buffer attwice the EC₅₀ concentration determined as described above isautomatically added by the FLIPR instrument to each well of the 96 wellplate for a final sample volume of 200 μl and a final capsaicinconcentration equal to the EC₅₀. The final concentration of testcompounds in the assay wells is between 1 μM and 5 μM. Antagonists ofthe capsaicin receptor decrease this response by at least about 20%,preferably by at least about 50%, and most preferably by at least 80%,as compared to matched control (i.e., cells treated with capsaicin attwice the EC₅₀ concentration in the absence of test compound), at aconcentration of 10 micromolar or less, preferably 1 micromolar or less.The concentration of antagonist required to provide a 50% decrease,relative to the response observed in the presence of capsaicin andwithout antagonist, is the IC₅₀ for the antagonist, and is preferablybelow 1 micromolar, 100 nanomolar, 10 nanomolar or 1 nanomolar.

Certain preferred VR1 modulators are antagonists that are essentiallyfree of agonist activity as demonstrated by the absence of detectableagonist activity in the assay described above at compound concentrationsbelow 4 nM, more preferably at concentrations below 10 μM and mostpreferably at concentrations less than or equal to 100 μM.

Example 7 Microsomal In Vitro Half-Life

This Example illustrates the evaluation of compound half-life values(t_(1/2) values) using a representative liver microsomal half-lifeassay.

Pooled human liver microsomes are obtained from XenoTech LLC (KansasCity, Kans.). Such liver microsomes may also be obtained from In VitroTechnologies (Baltimore, Md.) or Tissue Transformation Technologies(Edison, N.J.). Six test reactions are prepared, each containing 25 μlmicrosomes, 5 μl of a 100 μM solution of test compound, and 399 μl 0.1 Mphosphate buffer (19 mL 0.1 M NaH₂PO₄, 81 mL 0.1 M Na₂HPO₄, adjusted topH 7.4 with H₃PO₄). A seventh reaction is prepared as a positive controlcontaining 25 μl microsomes, 399 μl 0.1 M phosphate buffer, and 5 μl ofa 100 μM solution of a compound with known metabolic properties (e.g.,DIAZEPAM or CLOZAPINE). Reactions are preincubated at 39° C. for 10 min.

CoFactor Mixture is prepared by diluting 16.2 mg NADP and 45.4 mgGlucose-6-phosphate in 4 mL 100 mM MgCl₂. Glucose-6-phosphatedehydrogenase solution is prepared by diluting 214.3 μlglucose-6-phosphate dehydrogenase suspension (Roche MolecularBiochemicals; Indianapolis, Ind.) into 1285.7 μl distilled water. 71 μlStarting Reaction Mixture (3 mL CoFactor Mixture; 1.2 mLGlucose-6-phosphate dehydrogenase solution) is added to 5 of the 6 testreactions and to the positive control. 71 μl 100 mM MgCl₂ is added tothe sixth test reaction, which is used as a negative control. At eachtime point (0, 1, 3, 5, and 10 min), 75 μl of each reaction mix ispipetted into a well of a 96-well deep-well plate containing 75 μlice-cold acetonitrile. Samples are vortexed and centrifuged 10 min at3500 rpm (Sorval T 6000D centrifuge, H1000B rotor). 75 μl of supernatantfrom each reaction is transferred to a well of a 96-well platecontaining 150 μl of a 0.5 μM solution of a compound with a known LCMSprofile (internal standard) per well. LCMS analysis of each sample iscarried out and the amount of unmetabolized test compound is measured asAUC, compound concentration vs. time is plotted, and the t_(1/2) valueof the test compound is extrapolated.

Preferred compounds provided herein exhibit in vitro t_(1/2) values ofgreater than 10 min and less than 4 h, preferably between 30 min and 1h, in human liver microsomes.

Example 8 MDCK Toxicity Assay

This Example illustrates the evaluation of compound toxicity using aMadin Darby canine kidney (MDCK) cell cytotoxicity assay.

1 μL of test compound is added to each well of a clear bottom 96-wellplate (PACKARD, Meriden, Conn.) to give final concentration of compoundin the assay of 10 micromolar, 100 micromolar or 200 micromolar. Solventwithout test compound is added to control wells.

MDCK cells, ATCC no. CCL-34 (American Type Culture Collection, Manassas,Va.), are maintained in sterile conditions following the instructions inthe ATCC production information sheet. Confluent MDCK cells aretrypsinized, harvested, and diluted to a concentration of 0.1×10⁶cells/ml with warm (37° C.) medium (VITACELL Minimum Essential MediumEagle, ATCC catalog #30-2003). 100 μL of diluted cells is added to eachwell, except for five standard curve control wells that contain 100 μLof warm medium without cells. The plate is then incubated at 37° C.under 95% O₂, 5% CO₂ for 2 h with constant shaking. After incubation, 50μL of mammalian cell lysis solution (from the PACKARD (Meriden, Conn.)ATP-LITE-M Luminescent ATP detection kit) is added per well, the wellsare covered with PACKARD TOPSEAL stickers, and plates are shaken atapproximately 700 rpm on a suitable shaker for 2 min.

Compounds causing toxicity will decrease ATP production, relative tountreated cells. The ATP-LITE-M Luminescent ATP detection kit isgenerally used according to the manufacturer's instructions to measureATP production in treated and untreated MDCK cells. PACKARD ATP LITE-Mreagents are allowed to equilibrate to room temperature. Onceequilibrated, the lyophilized substrate solution is reconstituted in 5.5mL of substrate buffer solution (from kit). Lyophilized ATP standardsolution is reconstituted in deionized water to give a 10 mM stock. Forthe five control wells, 10 μL of serially diluted PACKARD standard isadded to each of the standard curve control wells to yield a finalconcentration in each subsequent well of 200 nM, 100 nM, 50 nM, 25 nMand 12.5 nM. PACKARD substrate solution (50 μL) is added to all wells,which are then covered, and the plates are shaken at approximately 700rpm on a suitable shaker for 2 min. A white PACKARD sticker is attachedto the bottom of each plate and samples are dark adapted by wrappingplates in foil and placing in the dark for 10 min. Luminescence is thenmeasured at 22° C. using a luminescence counter (e.g., PACKARD TOPCOUNTMicroplate Scintillation and Luminescence Counter or TECAN SPECTRAFLUORPLUS), and ATP levels calculated from the standard curve. ATP levels incells treated with test compound(s) are compared to the levelsdetermined for untreated cells. Cells treated with 10 μM of a preferredtest compound exhibit ATP levels that are at least 80%, preferably atleast 90%, of the untreated cells. When a 100 μM concentration of thetest compound is used, cells treated with preferred test compoundsexhibit ATP levels that are at least 50%, preferably at least 80%, ofthe ATP levels detected in untreated cells.

Example 9 Dorsal Root Ganglion Cell Assay

This Example illustrates a representative dorsal root ganglian cellassay for evaluating VR1 antagonist or agonist activity of a compound.

DRG are dissected from neonatal rats, dissociated and cultured usingstandard methods (Aguayo and White (1992) Brain Research 570:61-67).After 48 h incubation, cells are washed once and incubated for 30-60 minwith the calcium sensitive dye Fluo 4 AM (2.5-10 ug/ml; TefLabs, Austin,Tex.). Cells are then washed once. Addition of capsaicin to the cellsresults in a VR1-dependent increase in intracellular calcium levelswhich is monitored by a change in Fluo-4 fluorescence with afluorometer. Data are collected for 60-180 seconds to determine themaximum fluorescent signal.

For antagonist assays, various concentrations of compound are added tothe cells. Fluorescent signal is then plotted as a function of compoundconcentration to identify the concentration required to achieve a 50%inhibition of the capsaicin-activated response, or IC₅₀. Antagonists ofthe capsaicin receptor preferably have an IC₅₀ below 1 micromolar, 100nanomolar, 10 nanomolar or 1 nanomolar.

For agonist assays, various concentrations of compound are added to thecells without the addition of capsaicin. Compounds that are capsaicinreceptor agonists result in a VR1-dependent increase in intracellularcalcium levels which is monitored by a change in Fluo-4 fluorescencewith a fluorometer. The EC₅₀, or concentration required to achieve 50%of the maximum signal for a capsaicin-activated response, is preferablybelow 1 micromolar, below 100 nanomolar or below 10 nanomolar.

Example 10 Animal Models for Determining Pain Relief

This Example illustrates representative methods for assessing the degreeof pain relief provided by a compound.

A. Pain Relief Testing

The following methods may be used to assess pain relief.

Mechanical Allodynia

Mechanical allodynia (an abnormal response to an innocuous stimulus) isassessed essentially as described by Chaplan et al. (1994) J. Neurosci.Methods 53:55-63 and Tal and Eliav (1998) Pain 64(3):511-518. A seriesof von Frey filaments of varying rigidity (typically 8-14 filaments in aseries) are applied to the plantar surface of the hind paw with justenough force to bend the filament. The filaments are held in thisposition for no more than three seconds or until a positive allodynicresponse is displayed by the rat. A positive allodynic response consistsof lifting the affected paw followed immediately by licking or shakingof the paw. The order and frequency with which the individual filamentsare applied are determined by using Dixon up-down method. Testing isinitiated with the middle hair of the series with subsequent filamentsbeing applied in consecutive fashion, ascending or descending, dependingon whether a negative or positive response, respectively, is obtainedwith the initial filament.

Compounds are effective in reversing or preventing mechanicalallodynia-like symptoms if rats treated with such compounds requirestimulation with a Von Frey filament of higher rigidity strength toprovoke a positive allodynic response as compared to control untreatedor vehicle treated rats. Alternatively, or in addition, testing of ananimal in chronic pain may be done before and after compoundadministration. In such an assay, an effective compound results in anincrease in the rigidity of the filament needed to induce a responseafter treatment, as compared to the filament that induces a responsebefore treatment or in an animal that is also in chronic pain but isleft untreated or is treated with vehicle. Test compounds areadministered before or after onset of pain. When a test compound isadministered after pain onset, testing is performed 10 min to 3 h afteradministration.

Mechanical Hyperalgesia

Mechanical hyperalgesia (an exaggerated response to painful stimulus) istested essentially as described by Koch et al. (1996) Analgesia2(3):157-164. Rats are placed in individual compartments of a cage witha warmed, perforated metal floor. Hind paw withdrawal duration (i.e.,the amount of time for which the animal holds its paw up before placingit back on the floor) is measured after a mild pinprick to the plantarsurface of either hind paw.

Compounds produce a reduction in mechanical hyperalgesia if there is astatistically significant decrease in the duration of hindpawwithdrawal. Test compound may be administered before or after onset ofpain. For compounds administered after pain onset, testing is performed10 min to 3 h after administration.

Thermal Hyperalgesia

Thermal hyperalgesia (an exaggerated response to noxious thermalstimulus) is measured essentially as described by Hargreaves et al.(1988) Pain. 32(1):77-88. Briefly, a constant radiant heat source isapplied the animals' plantar surface of either hind paw. The time towithdrawal (i.e., the amount of time that heat is applied before theanimal moves its paw), otherwise described as thermal threshold orlatency, determines the animal's hind paw sensitivity to heat.

Compounds produce a reduction in thermal hyperalgesia if there is astatistically significant increase in the time to hindpaw withdrawal(i.e., the thermal threshold to response or latency is increased). Testcompound may be administered before or after onset of pain. Forcompounds administered after pain onset, testing is performed 10 min to3 h after administration.

B. Pain Models

Pain may be induced using any of the following methods, to allow testingof analgesic efficacy of a compound. In general, compounds providedherein result in a statistically significant reduction in pain asdetermined by at least one of the previously described testing methods,using male SD rats and at least one of the following models.

Acute Inflammatory Pain Model

Acute inflammatory pain is induced using the carrageenan modelessentially as described by Field et al. (1997) Br. J. Pharmacol.121(8):1513-1522. 100-200 μl of 1-2% carrageenan solution is injectedinto the rats' hind paw. Three to four hours following injection, theanimals' sensitivity to thermal and mechanical stimuli is tested usingthe methods described above. A test compound (0.01 to 50 mg/kg) isadministered to the animal, prior to testing, or prior to injection ofcarrageenan. The compound can be administered orally or through anyparenteral route, or topically on the paw. Compounds that relieve painin this model result in a statistically significant reduction inmechanical allodynia and/or thermal hyperalgesia.

Chronic Inflammatory Pain Model

Chronic inflammatory pain is induced using one of the followingprotocols:

-   -   1. Essentially as described by Bertorelli et al. (1999) Br. J.        Pharmacol. 128(6):1252-1258, and Stein et al. (1998) Pharmacol.        Biochem. Behay. 31(2):455-51, 200 μl Complete Freund's Adjuvant        (0.1 mg heat killed and dried M. Tuberculosis) is injected to        the rats' hind paw: 100 μl into the dorsal surface and 100 μl        into the plantar surface.    -   2. Essentially as described by Abbadie et al. (1994) J.        Neurosci. 14(10):5865-5871 rats are injected with 150 μl of CFA        (1.5 mg) in the tibio-tarsal joint.

Prior to injection with CFA in either protocol, an individual baselinesensitivity to mechanical and thermal stimulation of the animals' hindpaws is obtained for each experimental animal.

Following injection of CFA, rats are tested for thermal hyperalgesia,mechanical allodynia and mechanical hyperalgesia as described above. Toverify the development of symptoms, rats are tested on days 5, 6, and 7following CFA injection. On day 7, animals are treated with a testcompound, morphine or vehicle. An oral dose of morphine of 1-5 mg/kg issuitable as positive control. Typically, a dose of 0.01-50 mg/kg of testcompound is used. Compounds can be administered as a single bolus priorto testing or once or twice or three times daily, for several days priorto testing. Drugs are administered orally or through any parenteralroute, or applied topically to the animal.

Results are expressed as Percent Maximum Potential Efficacy (MPE). 0%MPE is defined as analgesic effect of vehicle, 100% MPE is defined as ananimal's return to pre-CFA baseline sensitivity. Compounds that relievepain in this model result in a MPE of at least 30%.

Chronic Neuropathic Pain Model

Chronic neuropathic pain is induced using the chronic constrictioninjury (CCl) to the rat's sciatic nerve essentially as described byBennett and Xie (1988) Pain 33:87-107. Rats are anesthetized (e.g. withan intraperitoneal dose of 50-65 mg/kg pentobarbital with additionaldoses administered as needed). The lateral aspect of each hind limb isshaved and disinfected. Using aseptic technique, an incision is made onthe lateral aspect of the hind limb at the mid thigh level. The bicepsfemoris is bluntly dissected and the sciatic nerve is exposed. On onehind limb of each animal, four loosely tied ligatures are made aroundthe sciatic nerve approximately 1-2 mm apart. On the other side thesciatic nerve is not ligated and is not manipulated. The muscle isclosed with continuous pattern and the skin is closed with wound clipsor sutures. Rats are assessed for mechanical allodynia, mechanicalhyperalgesia and thermal hyperalgesia as described above.

Compounds that relieve pain in this model result in a statisticallysignificant reduction in mechanical allodynia, mechanical hyperalgesiaand/or thermal hyperalgesia when administered (0.01-50 mg/kg, orally,parenterally or topically) immediately prior to testing as a singlebolus, or for several days: once or twice or three times daily prior totesting.

1. A compound of the formula:

or a pharmaceutically acceptable salt thereof, wherein: Ar₂ is phenyl ora 6-membered aromatic heterocycle, each of which is substituted withfrom 0 to 4 substituents independently chosen from R₂; X, Y and Z areindependently CR_(x) or N, such that at least one of X, Y and Z is N; D,K, J and F are independently N, CH or carbon substituted with asubstituent represented by R₁ or R₁₀; R_(x) is independently chosen ateach occurrence from hydrogen, halogen, C₁-C₄alkyl, amino, cyano andmono- or di-(C₁-C₄alkyl)amino; R₁ represents from 0 to 3 substituentsindependently chosen from: (a) halogen, cyano and nitro; and (b) groupsof the formula -Q-M-R_(y); R₁₀ represents one substituent chosen from:(a) —COOH, aminocarbonyl, imino, C₁-C₆alkoxycarbonyl or C₂-C₆alkanoyl;(b) C₁-C₆alkyl, C₂-C₆alkyl ether, C₁-C₆alkoxy, C₁-C₆alkylamino or mono-or di-(C₁-C₄alkyl)aminocarbonyl, each of which is substituted with from1 to 4 substituents independently chosen from: (i) halogen, hydroxy,—COON, cyano, amino and aminocarbonyl; and (ii) C₁-C₆alkoxy,C₃-C₈cycloalkyl, C₁-C₆alkylsulfonyl, C₁-C₆alkoxycarbonyl, C₂-C₄alkanoyl,mono- and di-(C₁-C₈alkyl)amino 4- to 7-membered heterocycles and phenyl,each of which is substituted with from 0 to 4 substituents independentlychosen from halogen, C₁-C₄alkyl and C₁-C₆alkoxy; or (c)(C₃-C₈cycloalkyl)aminocarbonyl or a group of the formula:

 wherein

 represents a 4- to 7-membered, N-linked heterocycloalkyl, each of whichis substituted with from 0 to 4 substituents independently chosen fromhydroxy, —COOH cyano, amino aminocarbonyl, C₁-C₆alkyl, C₂-C₆alkyl ether,C₃-C₈cycloalkyl, C₁-C₆alkoxy, C₁-C₆alkylsulfonyl, C₁-C₆alkoxycarbonyl,C₂-C₄alkanoyl, C₂-C₄alkanoylamino and mono- and di-(C₁-C₄alkyl)amino;such that R₁₀ is not hydroxy, amino or an unsubstituted group chosenfrom C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆alkoxy, C₂-C₆alkylether, C₂-C₆alkanoyl, C₃-C₆alkanone, C₁-C₆haloalkyl, C₁-C₆haloalkoxy,mono- or di-(C₁-C₆alkyl)amino, C₁-C₆alkylsulfonyl, mono- ordi-(C₁-C₆alkyl)aminosulfonyl or mono- or di-(C₁-C₆alkyl)aminocarbonyl;Each Q is independently chosen from C₀-C₄alkylene; Each M isindependently absent or selected from O, C(═O), OC(═O), C(═O)O,O—C(═O)(O, S(O)_(m), N(R_(z)), C(═O)N(R_(z)), C(═NH)N(R_(z)),N(R_(z))C(═O), N(R_(z))C(═NH), N(R_(z))S(O)_(m), S(O)_(m)N(R_(z)) andN[S(O)_(m)R_(z)]S(O)_(m); wherein m is independently selected at eachoccurrence from 0, 1 and 2; and R_(z) is independently selected at eachoccurrence from hydrogen, C₁-C₈alkyl and groups that are taken togetherwith R_(y) to form an optionally substituted 4- to 7-memberedheterocycle; and Each R_(y) is independently hydrogen, C₁-C₈haloalkyl,C₁-C₈alkyl, C₂-C₈alkenyl, (C₃-C₈carbocycle)C₀-C₄alkyl, (4- to 7-memberedheterocycle)C₀-C₄alkyl, or taken together with R_(z) to form a 4- to7-membered heterocycle, wherein each alkyl, carbocycle and heterocycleis substituted with from 0 to 4 substituents independently selected fromhydroxy, halogen, amino, cyano, nitro, oxo, —COOH, aminocarbonyl,aminosulfonyl, C₁-C₆alkyl, C₃-C₇cycloalkyl, C₂-C₆alkyl ether,C₁-C₆alkanoyl, C₁-C₆alkylsulfonyl, C₁-C₈alkoxy, C₁-C₈hydroxyalkyl,C₁-C₈alkylthio, mono- and di-(C₁-C₆alkyl)aminocarbonyl, mono- anddi-(C₁-C₆alkyl)aminosulfonyl, mono- and di-(C₁-C₆alkyl)amino,C₁-C₆alkanoylamino, and phenyl; such that R_(y) is not hydrogen if Q isC₀alkyl and M is absent; Each R₂ is: (a) independently chosen from (i)hydroxy, amino, cyano, halogen, —COOH, aminosulfonyl, nitro andaminocarbonyl; and (ii) C₁-C₆alkyl, (C₃-C₈cycloalkyl)C₀-C₄alkyl,C₁-C₆haloalkyl, C₁-C₆alkoxy, C₁-C₆alkylthio, C₂-C₆alkyl ether,C₂-C₆alkanoyl, C₁-C₆alkoxycarbonyl, C₂-C₆alkanoyloxy, C₃-C₆alkanone,mono- and di-(C₁-C₈alkyl)aminoC₀-C₆alkyl, (4- to 7-memberedheterocycle)C₀-C₄alkyl, C₁-C₆alkylsulfonyl, mono- anddi-(C₁-C₆alkyl)aminosulfonyl, and mono- anddi-(C₁-C₆alkyl)aminocarbonyl, each of which is substituted with from 0to 4 substituents independently chosen from halogen, hydroxy, cyano,amino, aminocarbonyl, aminosulfonyl, —COOH and oxo; or (b) takentogether with an adjacent R₂ to form a fused 5- to 13-memberedcarbocyclic or heterocyclic group that is substituted with from 0 to 3substituents independently chosen from halogen, oxo and C₁-C₆alkyl; R₃is selected from: (i) hydrogen and halogen; (ii) C₁-C₆alkyl,(C₃-C₈cycloalkyl)C₀-C₂alkyl, C₁-C₆haloalkyl and phenylC₀-C₂alkyl; and(iii) groups of the formula:

wherein: L is C₀-C₆alkylene or C₁-C₆alkylene that is taken together withR₅, R₆ or R₇ to form a 4- to 7-membered heterocycle; W is CO, S, SO orSO₂; R₅ and R₆ are: (a) independently chosen from hydrogen, C₁-C₁₂alkyl,C₂-C₁₂alkenyl, (C₃-C₈cycloalkyl)C₀-C₄alkyl, C₂-C₆alkanoyl,C₁-C₆alkylsulfonyl, phenylC₀-C₆alkyl, (4- to 7-memberedheterocycle)C₀-C₆alkyl and groups that are joined to L to form a 4- to7-membered heterocycle; or (b) joined to form a 4- to 12-memberedheterocycle; and R₇ is hydrogen, C₁-C₁₂alkyl, C₂-C₁₂alkenyl,(C₃-C₈cycloalkyl)C₀-C₄alkyl, C₂-C₆alkanoyl, phenylC₀-C₆alkyl, (4- to7-membered heterocycle)C₀-C₆alkyl or a group that is joined to L to forma 4- to 7-membered heterocycle; wherein each of (ii) and (iii) issubstituted with from 0 to 4 substituents independently chosen from: (1)halogen, hydroxy, amino, cyano, —COOH, aminosulfonyl, oxo, nitro andaminocarbonyl; and (2) C₁-C₆alkyl, (C₃-C₈cycloalkyl)C₀-C₂alkyl,C₁-C₆haloalkyl, C₁-C₆alkoxy, C₁-C₆alkanoyl, C₁-C₆alkoxycarbonyl,C₂-C₆alkanoylamino, mono- and di-(C₁-C₆alkyl)aminoC₀-C₄alkyl,C₁-C₆alkylsulfonyl, mono- and di-(C₁-C₆alkyl)aminosulfonyl, mono- anddi-(C₁-C₆alkyl)aminocarbonylC₀-C₄alkyl, phenylC₀-C₄alkyl and (4- to7-membered heterocycle)C₀-C₄alkyl, each of which is substituted withfrom 0 to 4 secondary substituents independently chosen from halogen,hydroxy, cyano, oxo, imino, C₁-C₄alkyl, C₁-C₄alkoxy and C₁-C₄haloalkyl;and R₄ represents from 0 to 2 substituents independently chosen fromC₁-C₃alkyl, C₁-C₃haloalkyl and oxo; wherein two of X, Y, and Z arenitrogen and the other is CR_(x).
 2. A compound or pharmaceuticallyacceptable salt according to claim 1, wherein the compound has theformula:


3. A compound or pharmaceutically acceptable salt according to claim 1,wherein D is N.
 4. A compound or pharmaceutically acceptable saltaccording to claim 1, wherein J, F and K are CH or carbon substitutedwith a substituent represented by R₁ or R₁₀.
 5. A compound orpharmaceutically acceptable salt according to claim 1, wherein onesubstituent represented by R₁ or R₁₀ is located meta or para to thepoint of attachment.
 6. A compound or pharmaceutically acceptable saltaccording to claim 1, wherein R₃ is a group of the formula:

wherein: L is C₀-C₆alkylene; and R₅ and R₆ are: (a) independently chosenfrom hydrogen, C₁-C₆alkyl, C₂-C₆alkenyl, (C₃-C₈cycloalkyl)C₀-C₄alkyl,C₂-C₄alkanoyl and groups that are joined to L to form a 4- to 7-memberedheterocycle; or (b) joined to form a 4- to 12-membered heterocycloalkyl;each of which alkyl, alkenyl, (cycloalkyl)alkyl, alkanoyl andheterocycloalkyl is substituted with from 0 to 4 substituentsindependently chosen from (i) halogen, hydroxy, amino, aminocarbonyl,oxo, —COOH and aminosulfonyl; and (ii) C₁-C₄alkyl, C₅-C₇cycloalkyl,C₁-C₄alkoxy, C₂-C₄alkanoyl, C₁-C₄haloalkyl, mono- anddi-(C₁-C₄alkyl)aminoC₀-C₂alkyl, mono- anddi-(C₁-C₄alkyl)aminocarbonylC₀-C₂alkyl, phenylC₀-C₄alkyl and (4- to7-membered heterocycle)C₀-C₂alkyl, each of which is substituted withfrom 0 to 4 secondary substituents independently chosen from halogen,hydroxy, cyano, C₁-C₄alkyl, C₁-C₄alkoxy and C₁-C₄haloalkyl.
 7. Acompound or pharmaceutically acceptable salt according to claim 6,wherein R₃ is azetidinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl,piperidinyl, piperazinyl, tetrahydropyridyl or azepanyl, each of whichis substituted with from 0 to 4 substituents independently chosen from:(a) halogen, hydroxy, amino, oxo, aminocarbonyl, aminosulfonyl and—COOH; and (b) C₁-C₄alkyl, C₂-C₄alkenyl, C₅-C₇cycloalkyl,C₁-C₄haloalkyl, C₁-C₄alkoxy, C₂-C₄alkyl ether, C₂-C₄alkanoyl,C₁-C₄alkylsulfonyl, C₂-C₄alkanoylamino and mono- anddi-(C₁-C₄alkyl)amino, each of which is substituted with from 0 to 4secondary substituents independently chosen from hydroxy and halogen. 8.A compound or pharmaceutically acceptable salt according to claim 1,wherein Ar₂ is phenyl, pyridyl or pyrimidyl, each of which issubstituted with from 1 to 3 substituents independently chosen fromamino, cyano, halogen, C₁-C₄alkyl, C₁-C₄haloalkyl, C₁-C₄aminoalkyl,C₁-C₄alkoxy, C₁-C₄haloalkoxy, C₁-C₄alkylthio, and mono- anddi-(C₁-C₄alkyl)aminoC₀-C₄alkyl.
 9. A compound or pharmaceuticallyacceptable salt according to claim 1, wherein the compound has theformula:

wherein: Ar₂ is phenyl, pyridyl or pyrimidyl, each of which issubstituted with from 0 to 3 substituents independently chosen fromamino, cyano, halogen, C₁-C₄alkyl, C₁-C₄haloalkyl, C₁-C₄aminoalkyl,C₁-C₄alkoxy, C₁-C₄haloalkoxy, C₁-C₄alkylthio, and mono- anddi-(C₁-C₄alkyl)aminoC₀-C₄alkyl; R_(1a) and R_(1b) are independentlyhydrogen, halogen, amino, cyano, C₁-C₆alkyl, C₁-C₆haloalkyl,C₁-C₆alkoxy, C₁-C₆alkylsulfonyl, mono- or di-(C₁-C₆alkyl)aminosulfonyl,(C₃-C₈cycloalkyl)aminocarbonyl or a group of the formula:

 wherein

 represents a 4- to 7-membered, N-linked heterocycloalkyl; R₄ represents0 substituents or one methyl substituent; and R₁₀ is: (a) —COOH,aminocarbonyl, imino, C₁-C₆alkoxycarbonyl or C₂-C₆alkanoyl; (b)C₁-C₆alkyl, C₂-C₆alkyl ether, C₁-C₆alkoxy, C₁-C₆alkylamino or mono- ordi-(C₁-C₄alkyl)aminocarbonyl, each of which is substituted with from 1to 4 substituents independently chosen from: (i) halogen, hydroxy,—COOH, cyano, amino and aminocarbonyl; and (ii) C₁-C₆alkoxy,C₃-C₈cycloalkyl, C₁-C₆alkylsulfonyl, C₁-C₆alkoxycarbonyl, C₂-C₄alkanoyl,mono- and di-(C₁-C₈alkyl)amino, 4- to 7-membered heterocycles andphenyl, each of which is substituted with from 0 to 4 substituentsindependently chosen from halogen, C₁-C₄alkyl and C₁-C₆alkoxy; or (c)(C₃-C₈cycloalkyl)aminocarbonyl or a group of the formula:

 wherein

 represents a 4- to 7-membered, N-linked heterocycloalkyl, each of whichis substituted with from 0 to 4 substituents independently chosen fromhydroxy, —COON, cyano, amino, aminocarbonyl, C₁-C₆alkyl, C₂-C₆alkylether, C₃-C₈cycloalkyl, C₁-C₆alkoxy, C₁-C₆alkylsulfonyl,C₁-C₆alkoxycarbonyl, C₂-C₄alkanoyl, C₂-C₄alkanoylamino and mono- anddi-(C₁-C₄alkyl)amino wherein one of X or Y is nitrogen and the otherCR_(x).
 10. A compound or pharmaceutically acceptable salt according toclaim
 1. wherein the group:


11. A compound or pharmaceutically acceptable salt according to claim 1,wherein R₃ is azetidinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl,piperidinyl, piperazinyl, tetrahydropyridyl or azepanyl, each of whichis substituted with from 0 to 4 substituents independently chosen from:(a) halogen, hydroxy, amino, oxo, aminocarbonyl, aminosulfonyl and—COOH; and (b) C₁-C₄alkyl, C₂-C₄alkenyl, C₅-C₇cycloalkyl,C₁-C₄haloalkyl, C₁-C₄alkoxy, C₂-C₄alkyl ether, C₂-C₄alkanoyl,C₁-C₄alkylsulfonyl, C₂-C₄alkanoylamino and mono- anddi-(C₁-C₄alkyl)amino, each of which is substituted with from 0 to 4secondary substituents independently chosen from hydroxy and halogen.12. A compound or pharmaceutically acceptable salt according to claim11, wherein Ar₂ is phenyl, pyridyl or pyrimidyl, each of which issubstituted with from 1 to 3 substituents independently chosen fromamino, cyano, halogen, C₁-C₄alkyl, C₁-C₄haloalkyl, C₁-C₄aminoalkyl,C₁-C₄alkoxy, C₁-C₄haloalkoxy, C₁-C₄alkylthio, and mono- anddi-(C₁-C₄alkyl)aminoC₀-C₄alkyl.
 13. A pharmaceutical composition,comprising at least one compound or pharmaceutically acceptable saltaccording to claim 1 in combination with a physiologically acceptablecarrier or excipient.
 14. A method for treating pain due to rheumatoidarthritis in a patient, comprising administering to a patient sufferingfrom pain due to rheumatoid arthritis a therapeutically effective amountof at least one compound or pharmaceutically acceptable salt accordingto claim 1 and thereby alleviating pain in the patient.
 15. A compoundor pharmaceutically acceptable salt according to claim 1, wherein thecompound is:4-{4-(3-Chloro-4-fluoro-phenyl)-6-[4-(3-nitro-pyridin-2-yl)-piperazin-1-yl]-pyrimidin-2-yl}-morpholine;4-(3-Chloro-4-fluoro-phenyl)-6-[4-(3-methyl-5-nitro-pyridin-2-yl)-piperazin-1-yl]-2-(2-methyl-pyrrolidin-1-yl)-pyrimidine;4-(3-Chloro-4-fluoro-phenyl)-6-[4-(3-methyl-5-nitro-pyridin-2-yl)-piperazin-1-yl]-2-(4-propyl-piperazin-1-yl)-pyrimidine;or4-(3-Chloro-4-fluoro-phenyl)-6-[4-(6-methoxy-3-nitro-pyridin-2-yl)-piperazin-1-yl]-2-(2-methyl-pyrrolidin-1-yl)-pyrimidine.